Fibrosis biomarker assay

ABSTRACT

Methods of diagnosis or of quantitation of fibrosis comprise conducting an immunoassay to measure neo-epitope containing protein fragments naturally present in a biofluid sample, and associating an elevation of said measure in said patient above a normal level with the presence or extent of fibrosis. The immunoassay is conducted by a method comprising: 
     contacting protein fragments naturally present in said sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to said immunological binding partner to measure therein protein fragments comprising said neo-epitope, and wherein said protein is collagen type III, collagen type I, collagen type IV, collagen type V, or collagen type VI, elastin, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, vimentin, or C-reactive protein.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of PCT/EP2008/064946 filed on Nov. 4, 2008, which claims Convention priority from GB0721713.6 filed in the United Kingdom on Nov. 5, 2007, GB0722748.1 filed in the United Kingdom on Nov. 20, 2007 and GB0802814.4 filed in the United Kingdom on Feb. 15, 2008, and also claims the benefit under 35 U.S.C. §1.119(e) of U.S. Provisional application No. 61/211,467 filed on Mar. 30, 2009 and U.S. Provisional application No. 61/289,081 filed on Dec. 22, 2009. The entire contents of each of the aforementioned patent applications are incorporated herein by this references.

INCORPORATION BY REFERENCE TO MATERIAL SUBMITTED ON A COMPACT DISC

A sequence listing file is submitted herewith.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to assays for biomarkers useful in the diagnosis of fibrosis disease and prognosis of its development, including biomarkers indicative of the risk of developing fibrosis after a chronic injury.

In particular, according to the present invention, biomarkers relating to degradation fragments of Collagen type I, III, IV, V, and VI, elastin, C-reactive protein, and proteoglycans including Biglycan, Decorin, Versican, and Perlecan are found to be useful.

2. Description of Related Art

Fibrotic diseases (including those listed in Table 1) are a leading cause of morbidity and mortality, e.g. cirrhosis with 800,000 death per year worldwide¹.

TABLE 1 Different fibrotic diseases² Tissue Examples of Causes Liver Viral hepatitis Schistosomiasis Steatohepatitis (Alcoholic or non-alcoholic) Lung Idiopathic pulmonary fibrosis (IPF) Systemic sclerosis (Scleroderma) Kidney Nephrogenic systemic fibrosis (NSF) Diabetes Untreated hypertension Heart Heart attack Hypertension Atherosclerosis Restenosis Eye Macular degeneration, retinal and vitreal retinopathy Skin Systemic sclerosis and scleroderma, keloids, hypertrophic scars, burns, genetic factors NFS Pancreas Autoimmune/hereditary causes Intestine Crohn's disease/inflammatory bowl disease Brain Alzheimer's disease, AIDS Bone marrow Cancer, ageing Multi-organ Surgical complications, chemotherapeutic drug-induced fibrosis, radiation- fibrosis induced fibrosis, mechanical injuries

A ‘fibrotic disease’ is any disease giving rise to fibrosis, whether as a main or a secondary symptom.

Fibrosis is the end result of chronic inflammatory reactions induced by a variety of stimuli including persistent infections, autoimmune reactions, allergic responses, chemical insults, radiation, and tissue injury. Fibrosis is characterized by the accumulation and reorganization of the extracellular matrix (ECM). Despite having obvious etiological and clinical distinctions, most chronic fibrotic disorders have in common a persistent irritant that sustains the production of growth factors, proteolytic enzymes, angiogenic factors, and fibrogenic cytokines, which together stimulate the deposition of connective tissue elements, especially collagens and proteoglycans, which progressively remodel and destroy normal tissue architecture^(3,4). Despite its enormous impact on human health, there are currently no approved treatments that directly target the mechanisms of fibrosis⁵.

The key cellular mediator of fibrosis is the myofibroblast, which when activated serves as the primary collagen-producing cell.

Extracellular Matrix (ECM)

Fibrogenesis is a dynamic process involving complex cellular and molecular mechanisms that usually originates from tissue injury⁶. Fibrogenesis is the result of an imbalance in normal ECM regulation that alters the concentration of macromolecules leading to increased tissue size and density, with progressively impaired function. These macromolecules are mainly fibrous proteins with structural and adhesive functions, such as collagens and proteoglycans.

Collagen

Collagens are widely distributed in the human body, i.e. ˜30% of the protein mass in the human body is composed of collagens. Collagens are responsible for the structural integrity of the ECM of most connective tissues. The ECM content results from a fine balance between synthesis and degradation tightly controlled through regulation of gene expression and protein secretion, but also through endogenous protease inhibition and protein degradation by metalloproteinases and cysteine proteases⁷⁻⁹. Table 2 lists the major collagen types with their major tissue distribution.

TABLE 2 Major collagen types and their tissue distribution. Collagen type Tissue distribution I Most connective tissues II Cartilage, vitreous humor III Extensible connective tissues, e.g. liver, skin, lung, vascular system IV Basement membranes V Tissues containing collagen I VI Most connective tissues VII Skin, bladder, oral mucosa, umbilical cord, amnion VIII Many tissues, especially endothelium XIII Endothelial cells, skin, eye, heart, skeletal muscle XIV Vessel, bone, skin, cartilage, eye, nerve, tendon, uterus XXI Vessel, heart, stomach, kidney, skeletal muscle, placenta

Type I collagen is the most abundant collagen and is found in most connective tissues. It is especially important for the structure of bone and skin where the major collagenous components are type I and III collagens¹⁰.

Collagen type I and III are the major components of liver and lung in a 1:1 ratio in healthy tissue. In addition, collagen type IV and VI are found in the basement membranes in most tissues. The most common localization of type V collagen is within the characteristic collagen fibrils, in association with the collagen type I and III¹⁰.

Some collagens have a restricted tissue distribution: for example, type II, which is found almost exclusively in cartilage¹¹.

During fibrogenesis the net amount of collagens increases¹²⁻¹⁴. Table 3 shows by way of example the collagen increase during liver fibrosis.

TABLE 3 Changes of the composition of collagen from normal to cirrhotic human liver ¹⁵. Collagen Collagen Distribution Distribution Collagen normal cirrhotic Times normal liver cirrhotic type Chains liver (mg/g) liver (mg/g) increased (%) liver (%) I α₁(I) α₂(I) 2 16 8 37 42 III α₁(III) 2 8 4 37 21 IV α₁(IV) 0.5 7 14 9 18 α₂(IV) V α₁(V) 0.9 7 8 17 18 α₂(V) α₃(V) VI α₁(VI) 0.01 0.1 10 0.2 0.3 α₂(VI)

Elastin

Elastin is a protein present in many connective tissues, primarily those that are elastic. It has a very high content of the amino acids glycine, valine, alanine, and proline, and has a molecular weight of 64 to 66 kDa. It is organised in an irregular or random coil conformation made up of 830 amino acids. Elastin is made by linking many soluble tropoelastin protein molecules, in a reaction catalyzed by lysyl oxidase, to make a massive insoluble, durable cross-linked array.

Elastin serves an important function in arteries as a medium for pressure wave propagation to help blood flow and is particularly abundant in large elastic blood vessels such as the aorta. Elastin is also very important in the lungs, elastic ligaments and the skin.

Despite much efforts devoted to the understanding of elastin synthesis and turnover, neo-epitopes originating from the proteolytic cleavage of this matrix molecules have until now not been associated with disease development in fibrosis.

Vimentin

Vimentin is a member of the intermediate filament family of proteins. Intermediate filaments are an important structural feature of eukaryotic cells. They, along with microtubules and actin microfilaments, make up the cytoskeleton. Although most intermediate filaments are stable structures, in fibroblasts, vimentin exists as a dynamic structure. This filament is used as a marker for mesodermally derived tissues, and as such has been used as an immunohistochemical marker for sarcomas.

Hertig and coworkers (Hertig et al., J Am Soc Nephrol. 2008 August; 19(8):1584-91) investigated if epithelial-to-mesenchymal transition in renal tubular epithelial cells of subjects with chronic allograft nephropathy could predict the progression of fibrosis in the allograft and measured vimentin expression in 83 biopsies from these. They did find an association between elevated vimentin expression and the intestinal fibrosis score at 1 year after surgery.

In another study of hepatic fibrosis, Meriden and colleagues (Meriden et al., Clin Gastro & Hepatol 2010; 8:289-296) found a significant association between vimentin expression (in biopsies obtained at F0 stage) and fibrosis progression, with elevated levels predicting rapid progression of the hepatic fibrosis. Accordingly, we wanted to investigate if circulating fragments of vimentin could serve as sensitive and specific biomarkers of fibrosis.

Proteoglycans

Proteoglycans are a diverse group of macromolecules, which covalently link a variable number of glycosaminoglycan (GAG) side chains to a core protein¹⁶. These GAGs are polymers of disaccharide repeats (e.g. N-acetyl glucosamine or N-acetyl galactosamine), which are acidic (negatively charged) due to hydroxyl, carboxylated and sulfated side groups on the disaccharide units. This makes them highly hydrophilic, thus aiding the diffusion of water and positive ions (e.g. sodium from extracellular fluids)¹⁷. Furthermore, GAGs have the ability to form non-covalent links with for example hyaluronic acid chains to form even larger molecular complexes¹⁶. Table 4 lists the most studied proteoglycans associated with connective tissue.

TABLE 4 Proteoglycans of the extracellular matrix of connective tissue Group Proteoglycans Origin Function Large extracellular Aggrecan ¹⁸ Articular cartilage Extracellular matrix stability proteoglycans (aggregating chondrocytes, intervertebral (hyaluronan binding) and hyaluronan-binding) disc, nasal cartilage Versican ^(19, 20) Connective tissue: fibroblast, Cell-cell and cell-matrix keratinocytes, smooth muscle interactions cells, mesangial cells Binding of sugars in Ca— dependent manner Neurocan ²¹ Nervous tissue Binds to neural cell adhesion molecules Brevican ²² Nervous tissue Extracellular matrix stability Small Leucine-rich Decorin ²³ Connective tissue, cartilage, Binds to and connect collagen proteoglycans (collagen- bone molecules (matrix stabilization binding) and thickness) Organogenesis Binding of TGFβ Biglycans ²⁴ Capillary endothelium, skin Cell differentiation (keratinocytes), epithelium of Binds and connect collagen kidney fibrils Fibromodulin Connective tissue, bone, Regulate orientation of collagen fibers ¹⁷ cartilage Lumican ²³ Cornea, muscle, cartilage, Controls spacing and kidney, lung, intestine thickness of collagen fibers Cell-associated Serglycins ²⁵ Widely distributed to Hemopoietic cell proteoglycans endothelium - intercellular differentiation compartments Adhesion and activation of lymphoid cells Syndecans ²⁶ Widely distributed - often cell Binds collagens, fibronectin, membrane bound thrombospondin, tenascin and bFGF Betaglycan ²⁷ Widely distributed TGFβ receptor and signaling Possible reservoir of TGFβ Basement membrane Perlecan ²⁸ All basement membranes Selective barrier for proteoglycans macromolecules Cell-adhesion

C-Reactive Protein

C-reactive protein (CRP) is an acute phase serum protein produced by the liver in response to different clinical conditions such as, inflammation, infection, or trauma²⁹. The production of CRP is induced by cytokines such as IL-6, released from the affected or damaged tissues. The physiological role of CRP is yet unknown and discussions on its pro- or anti-inflammatory actions are ongoing.

Proteases

The imbalance between synthesis and degradation of ECM during fibrogenesis, results from conversion of the low-density subendothelial matrix into matrix rich in interstitial collagens. The increase in collagen and proteoglycans may be due to one or both of (1) a decrease in protein production and (2) impaired protein degradation, and hence less matrix degradation. The decreased protein degradation has recently received increased attention. In the regulation of this process matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play important roles, as well as other proteases and their inhibitors, such as cystein proteases and the cystatins.

MMPs

MMPs are a large group of endopeptidases, capable of degrading most if not all components of the ECM. Presently, more than 25 MMPs have been found. MMPs are characterized by an active site containing a metal atom, typically zinc, and are secreted as zymogens. Different MMPs are expressed in different tissues. In Table 5 MMPs in the liver are shown.

TABLE 5 MMPs in the liver³⁰⁻³² Family Protease Source Substrate Collagenases MMP-1 HSC I, II, III, VII, VIII, X, gelatin MMP-8 Neutrophil I, II, III, V, VII, X, gelatin MMP-13 HSC, MFB, KC I, II, III, VII, X, gelatin Stromelysins MMP-3 HSC III, IV, V, IX, X, XI, gelatin, laminin, fibronectin, proteoglycans, glycoproteins, elastin, pro-MMP-1/13 MMP-10 HSC III, IV, V, gelatin, elastin, aggrecan MMP-11 HC PAI-1, week activity against matrix proteins Gelatinases MMP-2 HSC, MBF I, II, III, IV, V, VII, X, XI, gelagin, elastin, laminin MMP-9 KC, HSC, HC I, II, III, IV, V, VII, X, XI, gelagin, elastin, laminin MMP-7 HSC Entactin, gelatin, elastin, fibronectin, vitronectin, laminin, fibrinogen Metalloelastase MMP-12 Macrophages Elastin, gelatins, IV, laminin, fibronectin, entactin, vitronectin, proteoglycan, myelin basic protein, α1-antitripsin MT-MMPs MMP-14 HSC, MFB, KC I, II, III, gelatin, fibronectin, vitronectin, laminin, fibrinogen, MMP-15 HC, BDEC pro-MMP-2, pro-MMP-13 Pro-MMP-2, fibronectin, tenascin, laminin, aggrecan, perlecan

TIMPs block MMPs' proteolytic activity by binding in a substrate- and tissue-specific manner to MMP and membrane-type 1 metalloproteinase in a trimolecular complex (Table 6). During fibrosis TIMP levels increase dramatically, and MMP levels increase modestly or remain relatively static (except MMP-2) which in all gives a decrease in degradation of collagens.

TABLE 6 TIMPs in the liver³¹ Name Sources Metalloproteinase inhibited TIMP-1 HSC, MFB, KC, HC Pro-MMP-9, MMP-1, MMP-2, MMP-3, MMP-13 TIMP-2 KC, HSC MT-MMP-1, MT-MMP-2, proMMP-2, MMP-3, MMP-13, MMP-7 TIMP-3 HC MT-MMP-1, MT-MMP-2, TACE, MMP-13

Fibroblast Activation Protein

Fibroblast Activation Protein alpha subunit (FAPa or FAP, alpha) is an integral membrane gelatinase belonging to the serine protease family. FAPa is the alpha subunit and DPP4 (CD26) the beta subunit of a heterodimeric membrane-bound proteinase complex also known as 170 kDa Melanoma Membrane Gelatinase, Integral Membrane Serine Proteinase and Seprase. Some cells make only FAPa homodimers, some only DPP4 homodimers. The monomer is inactive. FAP, alpha is selectively expressed in reactive stromal fibroblasts of epithelial cancers, granulation tissue of healing wounds, and malignant cells of bone and soft tissue sarcomas³³. This protein is thought to be involved in the control of fibroblast growth or epithelial-mesenchymal interactions during development, tissue repair, and epithelial carcinogenesis. It has been shown that expression of FAP increase with the stage of fibrosis^(34,35).

Fibrosis Biomarkers

A number of biochemical markers have been suggested for fibrotic diseases, although not specific product of the disease. In Table 7 is an example of biochemical markers of liver fibrosis used in clinical trial. In addition there are a lot of examples of biomarkers of other fibrotic diseases^(12, 36-42).

TABLE 7 summarizes some of the known markers of liver fibrosis. Chronic liver Biomarker Parameters disease Reference One parameter CRP NASH ⁴³ Hyaluronan HCV ⁴⁴⁻⁴⁷ IGF-I HCV ⁴⁸ Leptin HCV ⁴⁹ PIIIP HCV ⁵⁰ Several parameters MP3 PIIINP, MMP1 HCV ^(51, 52) Zheng et al index HA, PIIICP, PIIINP, Laminin, C-IV Chronic ⁵³ hepatitis Lebensztjen et al Laminin-2, C-IV, MMP2, MMP9-TIMP1 index HBV ⁵⁴ index Tenascin, hyaluronana, Colalegn VI, TIMP-1 HBV ⁵⁵ Tsochatzis et al Leptin, adiponectin, resistin HCV, HBC, NASH ⁵⁶ index Patel et al index Hyaluronan, TIMP-1, α₂-macroglobulin HCV ⁵⁷ TIMP-1, tenascin, collagen IV, PIIINP, MMP2, NASH ⁵⁸ laminin, Hyaluronan Forns-index (76, 77) Age, platelet count, γGT, cholesterol HCV ^(51, 59-62) HIV/HCV FibroTest (76, 78) Haptoglobin, α₂-macroglobulin, apolipoprotein HCV ^(45, 51, 60, 61, 63-75) A1, γGT, bilirubin HIV/HCV NAFLD NAFLD in diabetes patients Actitest FibroTest + ALT HCV ^(65, 76-78) APRI (Wai-index) AST, platelet count HIV/HCV ^(45, 51, 60, 61, 64, 66, 79-87) HCV NAFLD Hepascore Bilirubin, γGT, hyaluronan, α₂-macroglobulin, age, HCV ^(51, 61, 64, 66, 88) gender HIV/HCV FIB-4 Platelet count, AST, ALT, age HIV/HCV ^(61, 83) SHASTA Hyaluronan, albumin, AST HIV/HCV ⁶¹ Fibroindex FORN + APRI HCV ⁸⁹ Fibrometer test Platelet count, prothrombin index, AST, α₂- HIV/HCV ^(51, 61, 64, 66, 81) macroglobulin, hyaluronan, urea, age HCV NAFLD NFSA Age, hyperglycaemia, body mass index, platelets, NAFLD ⁸¹ albumin, AST/ALT Ultrasound + APRI HCV ⁸² Metwally et al index Platelet count, albumin, AST, history of blood HCV ⁹⁰ transfusion, HBV core antibody Mohamadnejad et al Age, HBV DNA levels, alkaline phosphatase, HCV ⁹¹ index albumin, platelet counts, AST FibroSpect II Hyaluronan, TIMP-1, α₂-macroglobulin HCV ^(85, 92, 93) Stepwise Combination of APRI and Fibrotest HCV ⁹⁴ combination algorithms Imbert-Bismut index α₂ macroglobulin, AST, ALT γGT, total bilirubin, HCV ⁹⁵ albumin, α1 globulin, α₂ globulin, β globulin, γ globulin, apolipoprotein A₁ Nunes et al Age, Platelets, INR, CD4, AST/ALT, Hyaluronan, HCV/HIV ⁹⁶ YKL-40, PIIINP HCV Fibroscan +++ Fibroscan, Fibrotest, APRI, HCV ⁹⁷

U.S. Pat. No. 5,387,504 describes the neo-epitope VDIPEN released by the action of stromelysin at the aggrecan site N₃₄₁-F₃₄₂ and an RIA assay employing a monoclonal antibody specific for this neo-epitope. More generally the use of monospecific antibodies specific for fragments of aggrecan, generated by specific stromelysin cleavage are described. Elevations of stromelysin occur in osteoarthritis, rheumatoid arthritis, atherosclerotic lesions, gout, inflammatory bowel disease (IBD), idiopathic pulmonary fibrosis (IPF), certain cancers, joint injuries, and numerous inflammatory diseases. Stromelysin is reported to be elevated in idiopathic pulmonary fibrosis, and it is alleged that the assay can be conducted on blood or other biological fluids to detect stromelysin cleavage products of aggrecan and that quantitation of such fragments can be used diagnostically in respect of IPF as well as other conditions. However, no evidence for this is provided and there have to our knowledge been no subsequent publications validating this prediction. Such RIA assays have been commercially available for many years and no reports of their successful use in diagnosing or monitoring any fibrotic disease have appeared.

U.S. Pat. No. 7,225,080 discloses a method for diagnosis of an inflammatory, a fibrotic or a cancerous disease in a patient by measuring the values of at least four biochemical markers selected from the group consisting of α2-macroglobulin, AST (aspartate aminotransferase), ALT (alanine aminotransferase), GGT (gammaglutamyl transpeptidase), γ-globulin, total bilirubin, albumin, α1-globulin, α2-globulin, haptoglobin, β-globulin, apoA1, IL-10, TGF-β1, apoA2, and apoB in the serum or plasma of said patient, and subsequently combining said values in order to determine the presence of liver fibrosis and/or liver necroinflammatory lesions in said patient. The patent does not teach the quantitative measurement of peptide fragment carrying neo-epitopes generated during fibrotic disease.

U.S. Pat. No. 6,060,255 describes a method for diagnosing the degree of liver fibrosis, comprising the steps of measuring the concentration of type IV collagen high molecular weight form in a sample using an antibody that specifically binds to type IV collagen, and relating the measurement to the degree of liver fibrosis. Again, no use is made of neo-epitopes produced by proteolytic enzymes acting in the body. The sample is actually digested with pepsin, which may obscure the natural pattern of collagen cleavage in the sample.

U.S. Pat. No. 4,628,027 (Gay) discloses the production of antibodies specific for connective tissue proteins and, more particularly, the production of monoclonal antibodies by fused cell hybrids against human collagens and enzymes involved in collagen degradation. The use of monoclonal antibodies against connective tissue proteins to establish the collagen profile of histological, cytological and biological fluid samples is described. However, the patent does not describe the measurement of connective tissue proteins based on the binding of antibodies to neo-epitopes on said connective tissue proteins.

Guañabens N et al, J Bone Miner Res, 1998⁹⁸ evaluated the bone turnover markers N-telopeptide of type I collagen (NTX), C-telopeptide of type I collagen (CTX) and N-terminal pro-peptide of collagen type I (PINP) in patients with primary biliary cirrhosis, a disease with increased hepatic fibrosis. The level of NTX, CTX and PINP were elevated in patients compared to controls and correlated with the histological stage of the disease. The antibodies employed in the NTX were raised against a cathepsin K cleaved site in the N-terminal of collagen type I and are dependent on the neoepitope JYDGKGVG↓. The antibodies employed in the CTX were raised against a cathepsin K cleaved site in the C-terminal of collagen type I and are dependent on the neoepitope EKAHDGGR↓. These markers are located in telopeptides of collagen type I and not in the internal part (the triple helical part) of collagen type I. The monoclonal antibodies employed for the PINP assay were raised against an internal epitope in the PINP sequence which is not a neo-epitope.

Møller S et al, Gut., 1999⁹⁹ demonstrated that the C-terminal cross linked telopeptide of type I collagen (ICTP) was elevated in alcoholic cirrhosis patients compared to controls. The study described showed that a biochemical marker can reflect hepatic fibrosis. The ICTP polyclonal antibody has been raised against trypsin and collagenase cleaved collagen type I. However, the antibodies are not binding to a neo-epitope.

Rosen H N et al, Calcif Tissue Int, 2004¹⁰⁰ assessed the bone turnover markers N-telopeptide of type I collagen (NTX) and C-telopeptide of type I collagen (CTX) in women receiving hormone replacement treatment (HRT). In the study it was observed that the bone turnover markers decreased with treatment. The antibodies employed in the NTX were raised against a cathepsin K cleaved site in the N-terminal of collagen type I and are dependent on the neoepitope JYDGKGVG↓. The antibodies employed in the CTX were raised against a cathepsin K cleaved site in the C-terminal of collagen type I and are dependent on the neoepitope EKAHDGGR↓. In contrast to the present invention, these antibodies were used for evaluation of bone metabolism and not fibrosis.

Lein M et al, Eur Urol, 2007¹⁰¹ evaluated the use of the neo-epitope specific bone turnover markers N-telopeptide of type I collagen (NTX) and C-telopeptide of type I collagen (CTX) in prostate cancer patients receiving zoledronic acid. In the study it was observed that the bone turnover markers decreased with treatment. The antibodies employed in the NTX were raised against a cathepsin K cleaved site in the N-terminal of collagen type I and are dependent on the neoepitope JYDGKGVG↓. The antibodies employed in the CTX were raised against a cathepsin K cleaved site in the C-terminal of collagen type I and are dependent on the neoepitope EKAHDGGR↓. In contrast to the present invention, these antibodies were used for evaluation of the bone metabolism during invasion of bone metastases and not fibrosis.

PIIINP has been used in a number of studies to assess the severity of fibrotic disease¹⁰², in patients with skin fibrosis following severe burn trauma¹⁰³, for disease progression in noncirrhotic primary biliary cirrhosis¹⁰⁴, in primary biliary cirrhosis and chronic viral hepatitis C¹⁰⁵.

PIIINP and ICTP were measured in patients with fibrosis of the myocardium¹⁰⁶.

Many reports combine a set of biochemical markers to improve the predictive value of the biochemical index. Eleven different serum markers were measured in 205 patients with fibrotic staging from F0 to F4, and the most informative markers were alpha2 macroglobulin, alpha2 globulin (or haptoglobin), gamma globulin, apolipoprotein A1, gamma glutamyltranspeptidase, and total bilirubin¹⁰⁷. An index of these markers had a negative predictive value (100% certainty of absence of F2, F3, or F4) was obtained for scores ranging from zero to 0.10 (12% [41] of all patients), and high positive predictive value (>90% certainty of presence of F2, F3, or F4) for scores ranging from 0.60 to 1.00 (34% [115] of all patients).

However, in none of the above mentioned reports is it suggested that measurements of peptide fragments based on antibodies binding to neo-epitopes as now claimed might be useful for the assessment of patients with fibrotic disease.

BRIEF SUMMARY OF THE INVENTION

The present invention now provides a method of diagnosis of fibrosis comprising, conducting an immunoassay to measure neo-epitope containing protein fragments naturally present in a patient biofluid sample, and associating an elevation of said measure in said patient above a normal level with the presence of fibrosis, wherein said immunoassay is conducted by a method comprising:

contacting protein fragments naturally present in said sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to said immunological binding partner to measure therein protein fragments comprising said neo-epitope, and wherein said protein is collagen type I, collagen type III, collagen type IV, collagen type V or collagen type VI, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, CRP, or vimentin subject to the proviso that when the neo-epitopes are formed by cleavage of type I collagen, the cleavage is not at a site at which collagen type I is cleaved by cathepsin K. WO2009/059972 published on 14 May 2009 (after the priority date hereof) discloses assays for neo-epitopes of collagen III, but does not disclose that an elevated level of such a measure is to be associated with the presence or extent of fibrosis. Optionally, an assay according to this invention is based on one of the proteins named above other than collagen Type III or if based on collagen Type III utilises an immunological binding partner against one of the neoepitopes formed at the cleavage sites PGIPGRNGDP* SEQ ID NO1, *ESCPTGPQNY SEQ ID NO2, or PKGDTGPRGP* SEQ ID NO3 (where * marks the cleavage site). For these purposes, cardiovascular disease may not be regarded as fibrosis, or the fibrosis detected according to the invention may be other than fibrosis accompanying cardiovascular disease. Optionally, an elevated result in an immunoassay according to this invention is associated with skin fibrosis, lung fibrosis, or liver fibrosis.

The method may comprise the preliminary step of obtaining a patient biofluid sample.

The invention includes a method of immunoassay to measure neo-epitope containing protein fragments naturally present in body fluid sample, wherein said immunoassay is conducted by a method comprising:

contacting protein fragments naturally present in said sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to said immunological binding partner to measure therein protein fragments comprising said neo-epitope, and wherein said protein is neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, collagen type I, collagen type IV, collagen type V, collagen type VI, CRP, or vimentin subject to the proviso that when the neo-epitopes are formed by cleavage of type I collagen, the cleavage is not at a site at which collagen type I is cleaved by cathepsin K. Optionally, an assay according to this invention is based on one of the proteins named above other than collagen Type III or if based on collagen Type III utilises an immunological binding partner against one of the neoepitopes formed at the cleavage sites PGIPGRNGDP* SEQ ID NO1, *ESCPTGPQNY SEQ ID NO2, or PKGDTGPRGP* SEQ ID NO3 (where * marks the cleavage site).

Said immunological binding partner may have specific binding affinity for peptide fragments comprising a C-terminal neoepitope or an N-terminal neoepitope.

Specific reactivity with or immunological affinity for a neo-epitope will imply that the relevant immunological binding partner is not reactive with intact protein from which the neo-epitope derives. Preferably, said immunological binding partner is not reactive with a neo-epitope sequence, such as a sequence listed below, if the sequence is prolonged past the respective cleavage site.

The term ‘immunological binding partner’ as used herein includes polyclonal and monoclonal antibodies and also specific binding fragments of antibodies such as Fab or F(ab′)₂. Thus, said immunological binding partner may be a monoclonal antibody or a fragment of a monoclonal antibody having specific binding affinity.

Preferably, said peptide fragments are fragments of Type I, III, IV, V, or VI collagen, elastin, C-reactive protein, or one of the proteoglycans Biglycan, Decorin, Versican, and Perlecan. The connective tissue proteins are preferred. Preferably, the neo-epitope sequence to which the immunological binding partner binds is not found in any other protein or is not found in any of the other proteins to which the method of the invention relates.

Several candidate proteases may be responsible for the digestion of proteins in the fibrotic tissues. Most likely, this is the result of the large range of complicated processes resulting in different neo-epitope profiles dependent on the levels of disease.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

The invention will be further described and illustrated with reference to the following examples and the accompanying drawings, in which:

FIG. 1 shows a graph showing CO3 ELISA results of different biological samples: Pooled human serum samples (Serum); Human amniotic fluid (AF); Human fibroblast culture media (Fibr. Cltr.);

FIG. 2 a shows a graph showing CO3 serum levels in sham operated (s) and bile duct ligated rats at baseline (b) and at termination (t);

FIG. 2 b shows the corresponding delta-values of CO3 in rat serum: Termination levels-Baseline levels;

FIG. 3 shows a graph showing CO3 levels in different human serum samples. Normal serum: from healthy individuals. COPD: Chronic Obstructed Pulmonary Disease (leading to lung fibrosis). Scleroderma (leading to skin and lung fibrosis). HCV: Hepatitis virus C (leading to liver fibrosis);

FIG. 4 shows Liver weights and Liver scores determined in Example 5;

FIG. 5 shows levels of MMP-9 cleavage fragments of Type III collagen measured according to the invention in Example 5;

FIG. 6 shows levels of Type III collagen gene expression in BDL or sham-operated rats determined in Example 5;

FIG. 7 shows changes of expression levels of the MMP-9 cleavage fragment of Type III collagen reactive with the antibody used in Example 5 as determined by Western blot;

FIG. 8 shows the results of histology staining of liver sections obtained in Example 5;

FIG. 9 shows correlations between measurements of fragments of Type III collagen according to the invention with other liver biomarkers as determined in Example 5;

FIG. 10 shows results obtained on human serum samples in Example 6; and

FIG. 11 shows results obtained in testing the reactivity of a monoclonal antibody recognising an N-terminal neo-epitope from CRP.

FIG. 12 shows collagen accumulation in rat liver measured in Example 8.

FIG. 13 shows immunoassay results obtained in Example 8.

FIG. 14 shows the correlation of the immunoassay results of FIG. 13 with liver collagen content.

FIG. 15 shows a comparison of the results of an immunoassay according to the invention with measurements of hyaluronic acid and of Sirius red staining in Example 8.

FIG. 16 shows in the first panel the correlation of results from the immunoassay according to the invention with Sirius red staining and in the second panel the correlation between hyaluronic acid levels and Sirius red staining.

FIG. 17 shows the lack of correlation between the results of the immunoassay of the invention and hyaluronic acid levels.

FIG. 18 shows skin sections and skin thickness measurements described in Example 9.

FIG. 19 shows results from an immunoassay according to the invention in Example 9.

FIG. 20 shows in panel A Western Blot images obtained in Example 9 and in panel B corresponding immunoassay results according to the invention.

FIG. 21 shows a correlation between immunoassay results and skin thickness measurements.

FIG. 22 shows a correlation between urine immunoassay results and Western blot measurements described in Example 9.

DETAILED DESCRIPTION OF THE INVENTION Collagen Assays Collagen Type I

We have determined that the enzymes listed in the following table cleave type I collagen at least the following cleavage sites (marked “.”):

TABLE 8 Collagen type I cleavage sites. Protease Collagen type I MMP-2 V.PGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO4 MMP-2 S.VPGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO5 MMP-2 G.ISVPGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO6 MMP-9 G.ISVPGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO6 MMP-13 G.FQGPPGEPGEPGASGPMGPRGPPGPPG.K SEQ ID NO7 MMP-13 V.PGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO8 MMP-2 F.SGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRG.L SEQ ID NO9 MMP-9 F.SGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRG.L SEQ ID NO9 MMP-13 F.SGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRG.L SEQ ID NO9 MMP-9 G.LPGERGRPGAPGPAG.A SEQ ID NO10 MMP-13 G.LPGERGRPGAPGPAG.A SEQ ID NO10 MMP-2 G.LTGSPGSPGPDGKTGPPGPAG.Q SEQ ID NO11 MMP-2 E.RGSPGPAGPKGSPGEAGRPGEAGLPGAKG.L SEQ ID NO12 MMP-2 G.ERGSPGPAGPKGSPGEAGRPGEAGLPGAKG.L SEQ ID NO13 MMP-9 G.LTGSPGSPGPDGKTGPPGPAG.Q SEQ ID NO14 MMP-9 G.LTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPG.A SEQ ID NO15 MMP-9 G.LTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPGARG.Q SEQ ID NO16 MMP-13 G.LTGSPGSPGPDGKTGPPGPAG.Q SEQ ID NO14 MMP-13 G.ERGSPGPAGPKGSPGEAGRPGEAGLPGAKG.L SEQ ID NO13 MMP-9 G.QDGRPGPPGPPGARG.Q SEQ ID NO17 MMP-9 G.LTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPGARG.Q SEQ ID NO18 MMP-2 G.KDGEAGAQGPPGPAGPAGERGEQGPAGSPGF.Q SEQ ID NO19 MMP-2 G.ERGEQGPAGSPGF.Q SEQ ID NO20 MMP-3 E.RGVPGPPGAVGPAGKDGEAGAQGPPGPAGPAGERGEQGPAGSPGF.Q SEQ ID NO21 MMP-8 E.RGVPGPPGAVGPAGKDGEAGAQGPPGPAGPAGERGEQGPAGSPGF.Q SEQ ID NO21 — 113 PKGDTGPRGP.122 SEQ ID NO22

P indicates hydroxyproline, M indicates oxidised methionine, and K indicates hydroxylysine.

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type I collagen, excluding cleavage at a cathepsin K type I collagen site.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 9 N-terminal sequences of protease generated peptide fragments of Collagen type I. (The symbol “.” Indicates the cleavage site) Collagen I, alpha1 .ISVPGP SEQ ID NO23 .LPGPGG SEQ ID NO26 .ARGLPG SEQ ID NO29 .LPGERG SEQ ID NO32 .GGPPGP SEQ ID NO35 .ERGSPG SEQ ID NO38 .RGVPGP SEQ ID NO41 .RGEQGP SEQ ID NO44 .ARGAPG SEQ ID NO47 .PIGNVG SEQ ID NO50 .GADGPA SEQ ID NO53 .QRGVVG SEQ ID NO56 .PMGPPG SEQ ID NO59 .LQGPPG SEQ ID NO62 .FDFSF SEQ ID NO65 .AKGEAG SEQ ID NO68 .VQGPPG SEQ ID NO71 .RGSPGP SEQ ID NO74 .ARGQAG SEQ ID NO77 .KDGEAG SEQ ID NO80 .QGLPGP SEQ ID NO83 .AGLPGP SEQ ID NO86 .LAGPPG SEQ ID NO89 .PSGASG SEQ ID NO92 .AGQRGV SEQ ID NO95 .VVGLPG SEQ ID NO98 .TGDAGP SEQ ID NO175 .VPGPMG SEQ ID NO24 .FQGPPG SEQ ID NO27 .SGLDGA SEQ ID NO30 .VRGEPG SEQ ID NO33 .NSGEPG SEQ ID NO36 .LTGSPG SEQ ID NO39 .VGPAGK SEQ ID NO42 .PGERGV SEQ ID NO45 .PGDRGE SEQ ID NO48 .AAGRVG SEQ ID NO51 .GPQGIA SEQ ID NO54 .GLPGQR SEQ ID NO57 .MGPPGL SEQ ID NO60 .SAGAPG SEQ ID NO63 .DFSF SEQ ID NO66 .GIAGAP SEQ ID NO69 .LPGPPG SEQ ID NO72 .FAGPPG SEQ ID NO75 .NVGAPG SEQ ID NO78 .GEVGPP SEQ ID NO81 .IAGQRG SEQ ID NO84 .RGVVGL SEQ ID NO87 .EPGKQG SEQ ID NO90 .GKQGPS SEQ ID NO93 .ARGPAG SEQ ID NO96 .VGPPGP SEQ ID NO99 .PGPMGP SEQ ID NO25 .KNGDDG SEQ ID NO28 .LDGAKG SEQ ID NO31 .PGAKGA SEQ ID NO34 .DGVAGP SEQ ID NO37 .QDGRPG SEQ ID NO40 .ERGEQG SEQ ID NO43 .ANGAPG SEQ ID NO46 .AKGDAG SEQ ID NO49 .PPGPAG SEQ ID NO52 .GQRGVV SEQ ID NO55 .PGLPGP SEQ ID NO58 .DKGETG SEQ ID NO61 .RTGDAG SEQ ID NO64 .ATGAAG SEQ ID NO67 .IAGAPG SEQ ID NO70 .AGPKGS SEQ ID NO73 .QAGVMG SEQ ID NO76 .PAGERG SEQ ID NO79 .ARGERG SEQ ID NO82 .LTGPIG SEQ ID NO85 .AGPPGA SEQ ID NO88 .ATGFPG SEQ ID NO91 .GPPGPA SEQ ID NO94 .ASGPAG SEQ ID NO97 .GPPGPP SEQ ID NO100

Alternatively, suitable immunological binding partners may be specifically reactive with any of the following sequences at the C terminal of a peptide:

TABLE 10 C-terminal sequences of protease generated peptide fragments of Collagen type I (The symbol “.” Indicates the cleavage site). Collagen I, alpha1 QLSYGY. SEQ ID NO101 KGHRGF. SEQ ID NO104 APGPAG. SEQ ID NO107 GANGAP. SEQ ID NO110 EPGPVG. SEQ ID NO113 KGPAGE. SEQ ID NO116 PPGPPG. SEQ ID NO119 AVGPAG. SEQ ID NO122 APGPDG. SEQ ID NO125 KDGVRG. SEQ ID NO128 PGPAGF. SEQ ID NO131 SAGPPG. SEQ ID NO134 GEVGPP. SEQ ID NO81 PGPQGI. SEQ ID NO140 IAGQRG. SEQ ID NO84 GPSGEP. SEQ ID NO146 PVGPVG. SEQ ID NO149 EQGPSG. SEQ ID NO152 GPPGPP. SEQ ID NO100 QMGPRG. SEQ ID NO161 PGADGQ. SEQ ID NO164 PPGPKG. SEQ ID NO167 PKGPAG. SEQ ID NO170 GPAGRP. SEQ ID NO173 TGPRGP. SEQ ID NO177 EKSTGG. SEQ ID NO102 PSGPRG. SEQ ID NO105 FPGAVG. SEQ ID NO108 ANGAPG. SEQ ID NO46 EPGPTG. SEQ ID NO114 RGSPGP. SEQ ID NO74 PPGARG. SEQ ID NO120 PAGPAG. SEQ ID NO123 RGERGF. SEQ ID NO126 PAGPTG. SEQ ID NO129 EPGDAG. SEQ ID NO132 ATGFPG. SEQ ID NO91 GEKGSP. SEQ ID NO138 GPQGIA. SEQ ID NO54 GQRGVV. SEQ ID NO55 ERGPPG. SEQ ID NO147 PQGPRG. SEQ ID NO150 PRGPPG. SEQ ID NO153 GPPSAG. SEQ ID NO156 PPGPAG. SEQ ID NO52 PGPPGA. SEQ ID NO162 AGSPGF. SEQ ID NO165 PGERGA. SEQ ID NO168 GRNGDP. SEQ ID NO171 PPGPIG. SEQ ID NO174 PPGPQG. SEQ ID NO103 APGPQG. SEQ ID NO106 SEGPQG. SEQ ID NO109 SGPQGP. SEQ ID NO112 RGFPGA. SEQ ID NO115 LPGAKG. SEQ ID NO118 PGKAGE. SEQ ID NO121 AGPAGE. SEQ ID NO124 PAGPRG. SEQ ID NO127 TGARGA. SEQ ID NO130 PAGPPG. SEQ ID NO133 NAGPPG. SEQ ID NO136 GAPGTP. SEQ ID NO139 PQGIAG. SEQ ID NO142 RGPPGP. SEQ ID NO148 HRGFSG. SEQ ID NO151 PPGPRG. SEQ ID NO154 PPSAGF. SEQ ID NO157 TPGPQG. SEQ ID NO160 QGIAGQ. SEQ ID NO163 LPGPSG. SEQ ID NO166 PMGPPG. SEQ ID NO59 SPGEQG. SEQ ID NO172 TGDAGP. SEQ ID NO175

Collagen Type III

We have determined that the enzymes listed in the following table cleave type III collagen at least the following cleavage sites (marked *):

TABLE 11 Cleavage sites in collagen type III. Protease Neo-Epitope MMP-1 A*GIPGAPGLMGARGPPGPA*G SEQ ID NO178 MMP-1 K*GDPGPPGIPGRNGDPGI*P SEQ ID NO179 MMP-1 G*LAGPPGMPGPRGSPGPQG*V SEQ ID NO180 MMP-1 G*ERGLPGPPGIKGPAGIPGF*P SEQ ID NO181 MMP-1 G*IAGITGARGLAGPPGMPGPR*G SEQ ID NO182 MMP-1 G*IKGHRGFPGNPGAPGSPGPAG*Q SEQ ID NO183 MMP-1 A*RGLAGPPGMPGPRGSPGPQGV*K SEQ ID NO184 MMP-1 I*TGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO185 MMP-1 I*TGARGLAGPPGMPGPRGSPGPQGV*K SEQ ID NO186 MMP-1 G*ITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO187 MMP-1 G*VKGESGKPGANGLSGERGPPGPQG*L SEQ ID NO188 MMP-1 G*SRGAPGPQGPRGDKGETGERGAAG*I SEQ ID NO189 MMP-1 P*KGDAGQPGEKGSPGAQGPPGAPGPLG*I SEQ ID NO190 MMP-1 G*ITGARGLAGPPGMPGPRGSPGPQGV*K SEQ ID NO191 MMP-1 G*LRGGAGPPGPEGGKGAAGPPGPPGAAGTPG*L SEQ ID NO192 MMP-1 G*HAGAQGPPGPPGINGSPGGKGEMGPAGIPGAPG*L SEQ ID NO193 MMP-1 A*GKSGDRGESGPAGPAGAPGPAGSRGAPGPQGPRGDKGETGERGAAG*I SEQ ID NO194 MMP-1 G*LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG*L SEQ ID NO195 MMP-3 G*ERGLPGPPGIKGPAGIPGF*P SEQ ID NO196 MMP-3 A*VGGLAGYPGPAGPPGPPGPPGT*S SEQ ID NO197 MMP-3 K*DGTSGHPGPIGPPGPRGNRGER*G SEQ ID NO198 MMP-3 A*VGGLAGYPGPAGPPGPPGPPGTSGHPG*S SEQ ID NO199 MMP-3 G*IAGITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO200 MMP-3 A*PGAPGGKGDAGAPGERGPPGLAGAPGLRG*G SEQ ID NO201 MMP-3 A*VGGLAGYPGPAGPPGPPGPPGTSGHPGSPG*S SEQ ID NO202 MMP-2 A*IGSPGPAGPRGPVGPSGPPG*K SEQ ID NO203 MMP-3 + -8 G*AIGSPGPAGPRGPVGPSGPPG*K SEQ ID NO204 MMP-8 P*AGQQGAIGSPGPA*G SEQ ID NO205 MMP-8 G*GPPGVAGPPGGSGPAGPP*G SEQ ID NO206 MMP-8 L*AGPPGMPGPRGSPGPQG*V SEQ ID NO207 MMP-8 G*LSGERGPPGPQGLPGLA*G SEQ ID NO208 MMP-8 R*GLAGPPGMPGPRGSPGPQG*V SEQ ID NO209 MMP-8 G*LAGPPGMPGPRGSPGPQGV*K SEQ ID NO210 MMP-8 R*GLAGPPGMPGPRGSPGPQGV*K SEQ ID NO211 MMP-8 G*PQGPPGKNGETGPQGPPGP*T SEQ ID NO212 MMP-8 G*VKGERGSPGGPGAAGFPGAR*G SEQ ID NO213 MMP-8 A*RGLAGPPGMPGPRGSPGPQG*V SEQ ID NO214 MMP-8 N*GLSGERGPPGPQGLPGLAGTA*G SEQ ID NO215 MMP-8 A*VGGLAGYPGPAGPPGPPGPPGT*S SEQ ID NO216 MMP-8 G*SPGGKGEMGPAGIPGAPGLMGA*R SEQ ID NO217 MMP-8 T*GARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO218 MMP-8 V*KGESGKPGANGLSGERGPPGPQG*L SEQ ID NO219 MMP-8 G*VKGERGSPGGPGAAGFPGARGLPGPPGSNGNPGPPGPSGSPGKDGPPGPAG*N SEQ ID NO220 MMP-8 G*SPGAQGPPGAPGPLGIAGITGARGLAGPPG*M SEQ ID NO221 MMP-8 R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQ*G SEQ ID NO222 MMP-8 R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQ*G SEQ ID NO223 MMP-8 G*IAGITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO224 MMP-9 G*IKGPAGIPGFPG*M SEQ ID NO225 MMP-9 G*QPGVMGFPGPKG*N SEQ ID NO226 MMP-9 G*IKGPAGIPGFPGMK*G SEQ ID NO227 MMP-9 G*IKGPAGIPGFPGMKG*H SEQ ID NO228 MMP-9 I*PGAPGLMGARGPPGPAG*A SEQ ID NO229 MMP-9 G*ERGLPGPPGIKGPAGIP*G SEQ ID NO230 MMP-9 G*IPGAPGLMGARGPPGPAG*A SEQ ID NO231 MMP-9 G*FRGPAGPNGIPGEKGPAG*E SEQ ID NO232 MMP-9 P*GIPGQPGSPGSPGPPGIC*E SEQ ID NO233 MMP-9 G*ERGLPGPPGIKGPAGIPGF*P SEQ ID NO234 MMP-9 A*VGGLAGYPGPAGPPGPPGPPG*T SEQ ID NO235 MMP-9 G*VKGERGSPGGPGAAGFPGARG*L SEQ ID NO236 MMP-9 G*DAGAPGAPGGKGDAGAPGERGPPG*L SEQ ID NO237 MMP-9 Q*GPPGPTGPGGDKGDTGPPGPQGL*Q SEQ ID NO238 MMP-9 G*INGSPGGKGEMGPAGIPGAPGLM*G SEQ ID NO239 MMP-9 Q*GPPGEPGQAGPSGPPGPPGAIGPS*G SEQ ID NO240 MMP-9 P*GPPGINGSPGGKGEMGPAGIPGAP*G SEQ ID NO241 MMP-9 R*GLPGPPGSNGNPGPPGPSGSPGKDGPPGPAG*N SEQ ID NO242 MMP-9 G*KNGETGPQGPPGPTGPGGDKGDTGPPGPQG*L SEQ ID NO243 MMP-9 G*LPGIAGPRGSPGERGETGPPGPAGFPGAPG*Q SEQ ID NO244 MMP-9 G*INGSPGGKGEMGPAGIPGAPGLMGARGPPGPAG*A SEQ ID NO245 MMP-9 P*GINGSPGGKGEMGPAGIPGAPGLMGARGPPGPAG*A SEQ ID NO246 MMP-9 P*PGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG*L SEQ ID NO247 MMP-9 G*LKGENGLPGENGAPGPMGPRGAPGERGRPGLPGAAG*A SEQ ID NO248 MMP-9 G*NTGAPGSPGVSGPKGDAGQPGEKGSPGAQGPPGAPGPLG*I SEQ ID NO249 MMP-9 G*LMGARGPPGPAGANGAPGLRGGAGEPGKNGAKGEPGPRG*E SEQ ID NO250 MMP-9 G*LRGGAGPPGPEGGKGAAGPPGPPGAAGTPGLQGMPGERGGLGSPGPKG*D SEQ ID NO251 MMP-8 and -9 G*QQGAIGSPGPAGPRGPVGPSGPPG*K SEQ ID NO252 MMP-9 K*GDPGPPGIPGRNGDPGIPGQPG*S SEQ ID NO253 MMP-9 G*LRGGAGPPGPEGGKGAAGPPGPPG*A SEQ ID NO254 MMP-9 G*KNGETGPQGPPGPTGPGGDKGDTGPPGPQG*L SEQ ID NO255 MMP-9 G*YQGPPGEPGQAGPSGPPGPPG*A SEQ ID NO256 MMP-9 G*VAGPPGGSGPAGPPGPQG*V SEQ ID NO257 MMP-8, -9 and G*DKGEPGGPGADGVPGKDGPRGPTGPIGPPGPAG*Q SEQ ID NO258 -13 ADAMTS-5 Q*GHAGAQGPPGPPGIN*G SEQ ID NO259 CathepsinK A*GERGAPGPA*G SEQ ID NO260 CathepsinK A*GIPGFPGMK*G SEQ ID NO261 CathepsinK F*PGMKGHRGFD*G SEQ ID NO262 CathepsinK G*FPGARGLPGPPG*S SEQ ID NO263 CathepsinK A*GFPGARGLPGPPG*S SEQ ID NO264 CathepsinK P*PGPPGPPGTSGHP*G SEQ ID NO265 CathepsinK G*FPGMKGHRGFD*G SEQ ID NO266 CathepsinK Q*PGDKGEGGAPGLPGI*A SEQ ID NO267 CathepsinK R*GDKGETGERGAAGIK*G SEQ ID NO268 CathepsinK D*GRNGEKGETGAPGLK*G SEQ ID NO269 CathepsinK A*GQPGDKGEGGAPGLPGIA*G SEQ ID NO270 CathepsinK G*GPPGENGKPGEPGPKGD*A SEQ ID NO271 CathepsinK A*GIPGFPGMKGHRGFD*G SEQ ID NO272 CathepsinK R*GGAGEPGKNGAKGEPGPR*G SEQ ID NO273 CathepsinK K*GERGSPGGPGAAGFPGARGLPGPP*G SEQ ID NO274 CathepsinK I*PGVPGAKGEDGKDGSPGEPGANGLP*G SEQ ID NO275 CathepsinK G*AAGFPGARGLPGPPGSNGNPGPPGPS*G SEQ ID NO276 CathepsinK R*PGPPGPSGPRGQPGVMGFPGPKGN*D SEQ ID NO277 CathepsinK Q*GPPGPPGINGSPGGKGEMGPAGIPGAP*G SEQ ID NO278 CathepsinK A*GKDGESGRPGRPGERGLPGPPGIK*G SEQ ID NO279 CathepsinK A*GARGNDGARGSDGQPGPPGPPGTAGFPG*S SEQ ID NO280 CathepsinK S*PGVSGPKGDAGQPGEKGSPGAQGPPGAPG*P SEQ ID NO281 CathepsinK R*GSDGQPGPPGPPGTAGFPGSPGAKGEVGPA*G SEQ ID NO282 CathepsinK Q*GPPGPPGINGSPGGKGEMGPAGIPGAPGLM*G SEQ ID NO283 CathepsinK A*GPPGPPGPPGTSGHPGSPGSPGYQGPPGEPG*Q SEQ ID NO284 CathepsinK F*PGAPGQNGEPGGKGERGAPGEKGEGGPPGVA*G SEQ ID NO285 CathepsinK A*GFPGAPGQNGEPGGKGERGAPGEKGEGGPPG*V SEQ ID NO286 CathepsinK A*GARGNDGARGSDGQPGPPGPPGTAGFPGSPGAKGEVGPA*G SEQ ID NO287 CathepsinK R*GAAGEPGRDGVPGGPGMRGMPGSPGGPGSDGKPGPPGSQGESGRPGPPGPS*G SEQ ID NO288 CathepsinS G*IAGITGARGL*A SEQ ID NO289 CathepsinS A*GPPGPPGAAGTPGLQG*M SEQ ID NO290 CathepsinS N*GLSGERGPPGPQGLPG*L SEQ ID NO291 CathepsinS M*GARGPPGPAGANGAPGLR*G SEQ ID NO292 CathepsinS N*GLSGERGPPGPQGLPGLA*G SEQ ID NO293 CathepsinS G*IAGITGARGLAGPPGMPGPRG*S SEQ ID NO294 CathepsinS G*IAGITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO295 CathepsinS R*GGAGPPGPEGGKGAAGPPGPPGAAGTPGLQ*G SEQ ID NO296 CathepsinS S*GPKGDAGQPGEKGSPGAQGPPGAPGPLG*I SEQ ID NO297 CathepsinS G*IAGITGARGLAGPPGMPGPRGSPGPQGVK*G SEQ ID NO298 CathepsinS S*GPKGDAGQPGEKGSPGAQGPPGAPGPLG*I SEQ ID NO299 CathepsinS G*IAGITGARGLAGPPGMPGPRGSPGPQGVK*G SEQ ID NO300 CathepsinS A*VGGLAGYPGPAGPPGPPGPPGTSGHPGSPGSPGYQ*G SEQ ID NO301 CathepsinS E*PGPQGHAGAQGPPGPPGINGSPGGKGEMGPAGIPGAPG*L SEQ ID NO302 ADAMTS1 I*PGFPGMKGHR*G SEQ ID NO303 ADAMTS1 R*GSPGGPGAAGFPGAR*G SEQ ID NO304 ADAMTS1 K*GPAGIPGFPGMKGHR*G SEQ ID NO305 ADAMTS1 R*GLAGPPGMPGPRGSPGPQ*G SEQ ID NO306 ADAMTS1 A*GITGARGLAGPPGMPGPR*G SEQ ID NO307 ADAMTS1 L*GIAGITGARGLAGPPGMPGPR*G SEQ ID NO308 ADAMTS1 T*GARGLAGPPGMPGPRGSPGPQ*G SEQ ID NO309 ADAMTS1 Q*GPPGPPGINGSPGGKGEMGPAG*I SEQ ID NO310 ADAMTS1 L*PGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO311 ADAMTS1 A*GITGARGLAGPPGMPGPRGSPGPQ*G SEQ ID NO312 ADAMTS1 T*GARGLAGPPGMPGPRGSPGPQGVK*G SEQ ID NO313 ADAMTS1 R*GLPGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO314 ADAMTS1 G*RPGLPGAAGARGNDGARGSDGQPGPPG*P SEQ ID NO315 ADAMTS1 N*GAPGPMGPRGAPGERGRPGLPGAAGAR*G SEQ ID NO316 ADAMTS1 A*GSRGAPGPQGPRGDKGETGERGAAGIK*G SEQ ID NO317 ADAMTS1 R*GLAGPPGMPGPRGSPGPQGVKGESGKPGAN*G SEQ ID NO318 ADAMTS1 R*GLAGPPGMPGPRGSPGPQGVKGESGKPGANGL*S SEQ ID NO319 ADAMTS1 P*GPPGSNGNPGPPGPSGSPGKDGPPGPAGNTGAPGS*P SEQ ID NO320 ADAMTS1 T*GARGLAGPPGMPGPRGSPGPQGVKGESGKPGAN*G SEQ ID NO321 ADAMTS1 R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQGVKGER*G SEQ ID NO322 ADAMTS1 G*GPPGVAGPPGGSGPAGPPGPQGVKGERGSPGGPGAAGF*P SEQ ID NO323 ADAMTS1 K*SGDRGESGPAGPAGAPGPAGSRGAPGPQGPRGDKGETGERGAAGIK*G SEQ ID NO324 ADAMTS4 I*PGFPGMKGHR*G SEQ ID NO325 ADAMTS4 R*GLAGPPGMPGPR*G SEQ ID NO326 ADAMTS4 G*PQGLQGLPGTGGPP*G SEQ ID NO327 ADAMTS4 K*GPAGIPGFPGMKGHR*G SEQ ID NO328 ADAMTS4 R*GLAGPPGMPGPRGSPGPQG*V SEQ ID NO329 ADAMTS4 G*GPPGENGKPGEPGPKGDAGAP*G SEQ ID NO330 ADAMTS4 A*PGFRGPAGPNGIPGEKGPAGER*G SEQ ID NO331 ADAMTS4 E*KGSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO332 ADAMTS4 L*PGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO333 ADAMTS4 R*GAPGFRGPAGPNGIPGEKGPAGER*G SEQ ID NO334 ADAMTS4 R*GLPGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO335 ADAMTS4 R*GPVGPSGPPGKDGTSGHPGPIGPPGPR*G SEQ ID NO336 ADAMTS4 A*PGPQGPRGDKGETGERGAAGIKGHR*G SEQ ID NO337 ADAMTS4 R*GAPGPQGPRGDKGETGERGAAGIKGHR*G SEQ ID NO338 ADAMTS4 R*GFPGNPGAPGSPGPAGQQGAIGSPGPAGPR*G SEQ ID NO339 ADAMTS4 L*PGPPGIKGPAGIPGFPGMKGHRGFDGR*N SEQ ID NO340 ADAMTS4 D*AGQPGEKGSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO341 ADAMTS4 R*GPTGPIGPPGPAGQPGDKGEGGAPGLPGIAGPR*G SEQ ID NO342 ADAMTS4 K*GDAGQPGEKGSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO343 ADAMTS4 R*NGEKGETGAPGLKGENGLPGENGAPGPMGPR*G SEQ ID NO344 ADAMTS4 A*PGFRGPAGPNGIPGEKGPAGERGAPGPAGPRGA*A SEQ ID NO345 ADAMTS4 R*GAPGFRGPAGPNGIPGEKGPAGERGAPGPAGPR*G SEQ ID NO346 ADAMTS4 R*GSPGERGETGPPGPAGFPGAPGQNGEPGGKGER*G SEQ ID NO347 ADAMTS4 G*HAGAQGPPGPPGINGSPGGKGEMGPAGIPGAPGLMG*A SEQ ID NO348 ADAMTS4 R*GLAGPPGMPGPRGSPGPQGVKGESGKPGANGLSGER*G SEQ ID NO349 ADAMTS8 L*GIAGITGARGL*A SEQ ID NO350 ADAMTS8 I*PGFPGMKGHR*G SEQ ID NO351 ADAMTS8 R*GLAGPPGMPGPR*G SEQ ID NO352 ADAMTS8 Q*GPPGAPGPLGIAGITGAR*G SEQ ID NO353 ADAMTS8 A*GITGARGLAGPPGMPGPR*G SEQ ID NO354 ADAMTS8 A*GIPGAPGLMGARGPPGPAGAN*G SEQ ID NO355 ADAMTS8 R*GLAGPPGMPGPRGSPGPQGVKG*E SEQ ID NO356 ADAMTS8 K*GSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO357 ADAMTS8 L*PGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO358 ADAMTS8 K*DGTSGHPGPIGPPGPRGNRGER*G SEQ ID NO359 ADAMTS8 A*GITGARGLAGPPGMPGPRGSPGPQ*G SEQ ID NO360 ADAMTS8 R*GLAGPPGMPGPRGSPGPQGVKGESG*K SEQ ID NO361 ADAMTS8 R*GLAGPPGMPGPRGSPGPQGVKGESGKPGAN*G SEQ ID NO362 ADAMTS8 R*GLAGPPGMPGPRGSPGPQGVKGESGKPGANGL*S SEQ ID NO363 ADAMTS8 P*GPPGSNGNPGPPGPSGSPGKDGPPGPAGNTGAPGS*P SEQ ID NO364 ADAMTS8 R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQGVKGER*G SEQ ID NO365 ADAMTS8 K*SGDRGESGPAGPAGAPGPAGSRGAPGPQGPRGDKGETGERGA*A SEQ ID NO366 ADAMTS8 R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQGVKGERGSPGGPGAAGFPGAR*G SEQ ID NO367 MMP9 _*AIGPSG____*_ SEQ ID NO368 MMP9 117′ PGIPGRNGDP*. 124′ SEQ ID NO369 MMP9 142′ *ESCPTGPQNY 151′ SEQ ID NO370 MMP9 113′ PKGDTGPRGP*.′122 SEQ ID NO371

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type III collagen.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 12 N-terminal sequences of protease generated peptide fragments of Collagen type III. Collagen type III GIPGAP SEQ ID NO372 IAGITG SEQ ID NO375 TGARGL SEQ ID NO378 KGDAGQ SEQ ID NO381 GKSGDR SEQ ID NO383 DGTSGH SEQ ID NO135 GPPGVA SEQ ID NO158 GLSGER SEQ ID NO387 GARGLA SEQ ID NO111 GAPGEK SEQ ID NO141 IPGAPG SEQ ID NO117 GPPGPT SEQ ID NO391 GLPGPP SEQ ID NO394 GINGSP SEQ ID NO397 LMGARG SEQ ID NO400 DKGEPG SEQ ID NO403 PGMKGH SEQ ID NO406 FPGMKG SEQ ID NO409 GQPGDK SEQ ID NO412 GERGSP SEQ ID NO415 GPPGPP SEQ ID NO100 GFPGAP SEQ ID NO420 IAGITG SEQ ID NO375 GPPGSN SEQ ID NO423 PGPQGH SEQ ID NO426 SGDRGE SEQ ID NO429 PGPPGI SEQ ID NO432 GAPGFR SEQ ID NO435 GSPGER SEQ ID NO439 AIGPSG SEQ ID NO368 GAPGPQ SEQ ID NO445 GPTGPI SEQ ID NO448 SRGAPG SEQ ID NO451 AGQPGE SEQ ID NO454 AGQQGA SEQ ID NO457 GARGLA SEQ ID NO111 GSRGAP SEQ ID NO462 SPGAQG SEQ ID NO465 GIPGQP SEQ ID NO468 GDPGPP SEQ ID NO373 IKGHRG SEQ ID NO376 ITGARG SEQ ID NO379 LRGGAG SEQ ID NO382 LQGLPG SEQ ID NO384 VGGLAG SEQ ID NO155 AGPPGM SEQ ID NO145 GLAGPP SEQ ID NO388 KGESGK SEQ ID NO374 QPGVMG SEQ ID NO144 FRGPAG SEQ ID NO137 INGSPG SEQ ID NO392 KNGETG SEQ ID NO395 PGENGK SEQ ID NO398 YQGPPG SEQ ID NO401 GHAGAQ SEQ ID NO404 FPGARG SEQ ID NO407 PGDKGE SEQ ID NO410 GPPGEN SEQ ID NO413 PGVPGA SEQ ID NO416 GKDGES SEQ ID NO418 NTGAPG SEQ ID NO437 GLSGER SEQ ID NO387 GPKGDA SEQ ID NO424 PGFPGM SEQ ID NO427 GITGAR SEQ ID NO430 ESCPTG SEQ ID NO433 RPGLPG SEQ ID NO436 PQGLQG SEQ ID NO440 PGFRGP SEQ ID NO443 GFPGNP SEQ ID NO446 GDAGQP SEQ ID NO449 VAGPPG SEQ ID NO452 PGAPGG SEQ ID NO455 PGPPGP SEQ ID NO458 GRNGEK SEQ ID NO460 GGAGEP SEQ ID NO463 PGVSGP SEQ ID NO466 DAGAPG SEQ ID NO469 LAGPPG SEQ ID NO89 RGLAGP SEQ ID NO377 VKGESG SEQ ID NO380 ERGLPG SEQ ID NO385 AIGSPG SEQ ID NO143 LSGERG SEQ ID NO176 IGSPGP SEQ ID NO386 PQGPPG SEQ ID NO389 VKGERG SEQ ID NO159 IKGPAG SEQ ID NO169 QQGAIG SEQ ID NO390 GPPGEP SEQ ID NO393 LPGIAG SEQ ID NO396 LKGENG SEQ ID NO399 GERGAP SEQ ID NO402 GSDGQP SEQ ID NO405 GFPGAR SEQ ID NO408 GDKGET SEQ ID NO411 GIPGFP SEQ ID NO414 AAGFPG SEQ ID NO417 GARGND SEQ ID NO419 GAAGEP SEQ ID NO421 GARGPP SEQ ID NO422 GGAGPP SEQ ID NO425 GSPGGP SEQ ID NO428 GIAGIT SEQ ID NO431 HAGAQG SEQ ID NO434 GAPGPM SEQ ID NO438 GPAGIP SEQ ID NO441 KGSPGA SEQ ID NO444 GPVGPS SEQ ID NO447 NGEKGE SEQ ID NO450 PGPQGP SEQ ID NO453 PGAPGQ SEQ ID NO456 SPGGKG SEQ ID NO459 GPPGAP SEQ ID NO461 GSPGAQ SEQ ID NO464 PGAPGL SEQ ID NO467 GPPGIN SEQ ID NO470 or with any of the following sequences at the C-terminal of a peptide:

TABLE 13 C-terminal sequences of protease generated peptide fragments of Collagen type III. Collagen type III GPPGPA SEQ ID NO94 GMPGPR SEQ ID NO473 ERGAAG SEQ ID NO476 ERGPPG SEQ ID NO147 PSGPPG SEQ ID NO483 GLPGLA SEQ ID NO486 GLAGTA SEQ ID NO488 LAGPPG SEQ ID NO89 IPGFPG SEQ ID NO492 FPGAPG SEQ ID NO494 GPPGIC SEQ ID NO2187 PGPQGL SEQ ID NO497 SPGPKG SEQ ID NO499 TGAPGS SEQ ID NO502 PGPKGD SEQ ID NO506 GLPGIA SEQ ID NO507 GLPGPP SEQ ID NO394 IPGAPG SEQ ID NO117 GEVGPA SEQ ID NO514 GEVGPA SEQ ID NO499 EKGPAG SEQ ID NO515 TSGHPG SEQ ID NO518 GTPGLQ SEQ ID NO521 GPQGVK SEQ ID NO524 PPGPAG SEQ ID NO52 FPGMKG SEQ ID NO409 GLSGER SEQ ID NO387 GMKGHR SEQ ID NO531 EMGPAG SEQ ID NO534 GRNGDP SEQ ID NO171 GVKGER SEQ ID NO538 PQGVKG SEQ ID NO541 GPPGPR SEQ ID NO544 AGPRGA SEQ ID NO547 GPAGAN SEQ ID NO550 NGDPGI SEQ ID NO471 SPGPAG SEQ ID NO474 PGPLGI SEQ ID NO477 PGPPGT SEQ ID NO479 APGLRG SEQ ID NO481 GSPGPA SEQ ID NO484 GPPGPQ SEQ ID NO490 FPGPKG SEQ ID NO491 GPAGIP SEQ ID NO441 PPGPPG SEQ ID NO119 GAIGPS SEQ ID NO495 LPGAAG SEQ ID NO2188 GAPGLM SEQ ID NO498 GEPGPR SEQ ID NO500 GHRGFD SEQ ID NO503 PGLPGI SEQ ID NO504 PQGLPG SEQ ID NO508 GANGLP SEQ ID NO510 GPPGIK SEQ ID NO512 PPGPQG SEQ ID NO103 GFPGAR SEQ ID NO408 EPGPRG SEQ ID NO516 GAPGPA SEQ ID NO519 GTSGHP SEQ ID NO522 GAPGLK SEQ ID NO525 PGPKGN SEQ ID NO527 PGANGL SEQ ID NO529 TGPRGP SEQ ID NO177 EGGPPG SEQ ID NO532 GIAGPR SEQ ID NO535 GKPGAN SEQ ID NO537 PGAAGF SEQ ID NO539 GDAGAP SEQ ID NO542 GPAGPR SEQ ID NO545 GGKGER SEQ ID NO548 SPGPQG SEQ ID NO472 PGPQGV SEQ ID NO475 AAGTPG SEQ ID NO478 GNRGER SEQ ID NO480 HPGSPG SEQ ID NO482 GPAGPP SEQ ID NO485 QGPPGP SEQ ID NO487 PGLMGA SEQ ID NO489 FPGARG SEQ ID NO407 GFPGMK SEQ ID NO493 AGIPGF SEQ ID NO496 APGPLG SEQ ID NO2189 GPPGIN SEQ ID NO470 IPGQPG SEQ ID NO501 LPGPPG SEQ ID NO72 GAAGIK SEQ ID NO505 GAPGLR SEQ ID NO509 GPPGPS SEQ ID NO511 TAGFPG SEQ ID NO513 GPPGVA SEQ ID NO158 TGARGL SEQ ID NO378 PPGAPG SEQ ID NO517 TPGLQG SEQ ID NO520 MPGPRG SEQ ID NO523 GSPGYQ SEQ ID NO526 GAAGAR SEQ ID NO528 GTGGPP SEQ ID NO530 GITGAR SEQ ID NO430 GSPGPQ SEQ ID NO533 QPGPPG SEQ ID NO536 VKGESG SEQ ID NO380 TGERGA SEQ ID NO540 GPAGER SEQ ID NO543 RGFDGR SEQ ID NO546 APGLMG SEQ ID NO549

Collagen IV

We have determined that the enzymes listed in the following table cleave type IV collagen at least the following cleavage sites (marked “.”):

TABLE 14 Cleavage fragments of collagen type IV Protease Neo-Epitope FAP D.IDGYRGPPGP.Q SEQ ID NO551 FAP S.MGPPGTPSVDHGF.L SEQ ID NO552 FAP P.DGLPGSMGPPGTPSVDHG.F SEQ ID NO553 FAP P.DGLPGSMGPPGTPSVDHGF.L SEQ ID NO554 FAP P.DGLPGSMGPPGTPSVDHGFL.V SEQ ID NO555 FAP P.SGRDGLPGPPGSPGPPGQPGY.T SEQ ID NO556 FAP P.SGRDGLPGPPGSPGPPGQPGYTN.G SEQ ID NO557 FAP P.SGRDGLPGPPGSPGPPGQPGYTNG.I SEQ ID NO558 FAP I.PGSKGEQGFMGPPGPQGQPGLPGS.P SEQ ID NO559 FAP P.RGFPGPPGPDGLPGSMGPPGTPSVD.H SEQ ID NO560 FAP E.PGPPGLPGSVGSPG.V SEQ ID NO561 FAP I.DGYRGPPGPQGP.P SEQ ID NO562 FAP P.RGFPGPPGPDGLPGSMG.P SEQ ID NO563 FAP D.GLPGSMGPPGTPSVDHGF.L SEQ ID NO564 FAP D.GLPGSMGPPGTPSVDHGFL.V SEQ ID NO565 FAP P.GLPGQQGAPGIPGFPGSKGEMGVMGTP.G SEQ ID NO566 FAP I.GIPGMPGSPGLKGSPGSVGYPGSPGLPGE.K SEQ ID NO567 FAP P.GPPGPPGEKGQMGLSFQGPKGDKGDQGVSGPPGVP.G SEQ ID NO568 FAP P.GIGPPGARGPPGGQGPPGLSGPPGIKGEKGFPGFPGL.D SEQ ID NO569 FAP E.PGLPGIPGVSGPK.G SEQ ID NO570 FAP G.EKGQKGDTGPPGPPGLV.I SEQ ID NO571 FAP L.PGIGVQGPPGPPGIPGPIGQPGLHGIPGEKGDPGPP.G SEQ ID NO572 FAP G.SPGIPGHQGEMG.P SEQ ID NO573 FAP E.PGMQGEPGPPGP.P SEQ ID NO574 FAP G.PPGRLGAPGTPGLPGP.R SEQ ID NO575 FAP P.PGPKGFPGIPGP.P SEQ ID NO576 FAP A.KGQPGLPGFPGT.P SEQ ID NO577 FAP D.RGPPGPPGIRGPPGP.P SEQ ID NO578 FAP P.GPPGEKGKPGQDGIPGP.A SEQ ID NO579 FAP L.LGSKGEKGEPGLPGIPGVSGPKGY.Q SEQ ID NO580 MMP-9 D.GLPGSMGPPGTPSVDHG.F SEQ ID NO581 MMP-9 D.GLPGSMGPPGTPSVDHGF.L SEQ ID NO564 MMP-9 T.GPLGEKGERGYPGTPGPRGE.P SEQ ID NO582 MMP-9 G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L SEQ ID NO583 MMP-9 P.DGLPGSMGPPGTPSVDHGFL.V SEQ ID NO555 MMP-9 D.PGLKGDKGDVGLPGKPGSMDKVDMGS.M SEQ ID NO584 MMP-9 L.PGPMGPPGLPGIDGV.K SEQ ID NO585 MMP-9 D.GLPGSMGPPGTPSVDHGFL.V SEQ ID NO565 MMP-9 G.IRGEPGPPGLPGSVGSPGVPGIGPPG.A SEQ ID NO586 MMP-9 G.FPGPPGPDGLPGSMGPPGTPSVDHGF.L SEQ ID NO587 MMP-9 G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPG.A SEQ ID NO588 MMP-9 G.IRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L SEQ ID NO589 MMP-9 E.DGVIGMMGFPGAIGP.P SEQ ID NO590 MMP-9 Y.PGNPGILGPPGEDGVIGMMGFPGAIGPPGPPG.N SEQ ID NO591 MMP-9 I.PPSDEICEPGPPGP.P SEQ ID NO592 MMP-9 L.PGLPGPKGEPGLPGYPGNPGIKGS.V SEQ ID NO593 MMP-9 G.IKGDKGSMGHPGPKGPP.G SEQ ID NO594 MMP-9 T.PGSPGCAGSPGLPGSPGPPG.P SEQ ID NO595 MMP-9 P.GAPGPQGLPGPPGFPGPVGPPGPPGFFGFPGAMGPRGPKGHMGE.R SEQ ID NO596 MMP-9 G.LPGFAGNPGP SEQ ID NO597 MMP-9 + FAP G.AEGLPGSPGFPGPQG.D SEQ ID NO598 MMP-9 + FAP M.GPPGVPGFQGPKGLP.G SEQ ID NO599 MMP-9 + FAP D.IDGYRGPPGPQGPPG.E SEQ ID NO600 MMP-9 + FAP G.DQGDQGVPGAKGLPGP.P SEQ ID NO601 MMP-9 + FAP G.DRGPQGQPGLPGLPGP.M SEQ ID NO602 MMP-9 + FAP P.DGLPGSMGPPGTPSVDHGF.L SEQ ID NO554 MMP-9 + FAP E.KGSIGIPGMPGSPGLKGSPGSVGYP.G SEQ ID NO603 MMP-9 + FAP G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPG.A SEQ ID NO588 MMP-9 + FAP G.IRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L SEQ ID NO589 MMP-9 + FAP G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L SEQ ID NO583 MMP-9 + FAP G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPGLSGPPG.I SEQ ID NO604 MMP-9 + FAP I.PPSDEICEPGPPGP.P SEQ ID NO592 MMP-9 + FAP P.GPPGLMGPPGPPGLPGP.K SEQ ID NO605 MMP-9 + FAP G.ERGSPGIPGAPGPIGPPGSPG.L SEQ ID NO606 MMP-9 + FAP P.GIPGAPGAPGFPGSKGEPGDILTFPGMKGDKGELGSPGAPGL.P SEQ ID NO607 MMP-9 + FAP C.DGGVPNTGPPGEPGPP.G SEQ ID NO608 MMP12, Alpha1 .ILGHVPGML. SEQ ID NO2190 MMP12, Alpha1 .PGLPGQPGPPGLPVPGQ. SEQ ID NO2191 MMP12, Alpha1 .SGYPGNPGLPGIPGQDGPPGPPGIPGCNGTKGERGPLGPPGL. SEQ ID NO2192 MMP12, Alpha1 .VSGPPGVPGQA. SEQ ID NO2193 MMP12, Alpha1 .VSGPPGVPGQAQ. SEQ ID NO2194 MMP12, Alpha2 .KRGPPGPPGLPGPPGPDGFL. SEQ ID NO2195 MMP12, Alpha2 .LHGFPGAPGQEGPLG. SEQ ID NO2196 MMP12, Alpha2 .LPGPDGPPGERGLPGEVL. SEQ ID NO2197 MMP12, Alpha2 .LRGIPGF. SEQ ID NO2198 MMP12, Alpha2 .PGFPGAPGTVGAPGIAGIPQK. SEQ ID NO2199 MMP12, Alpha2 .QQGNRGLGF. SEQ ID NO2200 MMP12, Alpha2 .VGQPGPNGIPSDTL. SEQ ID NO2201 MMP12, Alpha3 .GEPGMQGEPGPPGPPGNLGPCGPRGKPGKDGKPGTPGPAGEKG. SEQ ID NO2202 MMP12, Alpha3 .GEPGPPGPPGNLGPCGPRGKPGKDGKPGTPGPAGEKGNK. SEQ ID NO2203 MMP12, Alpha3 .PGIPGTPGPPGLPGLQGPVGPPG. SEQ ID NO2204 MMP12, Alpha3 .PGDIVFRK. SEQ ID NO2205 MMP12, Alpha4 .GNKGDPASHFGPPGPKG. SEQ ID NO2206 MMP12, Alpha4 .PGPRGKPGM. SEQ ID NO2207 MMP12, Alpha5 .PGLPGQPGTRGL. SEQ ID NO2208 MMP12, Alpha5 .PGPPGPLGIPGRSGVPGLKGDDGLQGQPGLPGPTGEKGSK. SEQ ID NO2209 MMP12, Alpha5 .PGPPGPLGIPGRSGVPGLKGDDGLQGQPGLPGPTGEKGSKG. SEQ ID NO2210 MMP12, Alpha5 .SKGEKGEPGLPGIPGVSGPKGYQGLPGDPGQPGLSGQPGL. SEQ ID NO2211

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type IV collagen.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 15 N-terminal sequences of protease generated peptide fragments of Collagen type IV. Collagen type IV IDGYRG SEQ ID NO609 PGSKGE SEQ ID NO612 GPPGPP SEQ ID NO100 LGSKGE SEQ ID NO617 PGIGVQ SEQ ID NO618 PGPKGF SEQ ID NO621 PPSDEI SEQ ID NO623 IRGEPG SEQ ID NO626 DGVIGM SEQ ID NO629 IKGDKG SEQ ID NO631 GPPGVP SEQ ID NO633 KGSIGI SEQ ID NO635 ERGSPG SEQ ID NO38 SGRDGL SEQ ID NO639 DGYRGP SEQ ID NO642 GIPGMP SEQ ID NO645 .ILGHVP SEQ ID NO2212 .PGLPGQ SEQ ID NO2213 .SGYPGN SEQ ID NO2217 .VSGPPG SEQ ID NO2220 .KRGPPG SEQ ID NO2223 .LHGFPG SEQ ID NO2225 MGPPGT SEQ ID NO610 RGFPGP SEQ ID NO613 GLPGSM SEQ ID NO615 GIGPPG SEQ ID NO611 SPGIPG SEQ ID NO619 KGQPGL SEQ ID NO622 PGLKGD SEQ ID NO624 FPGPPG SEQ ID NO627 PGNPGI SEQ ID NO630 PGSPGC SEQ ID NO632 DQGDQG SEQ ID NO634 PPGRLG SEQ ID NO636 GIPGAP SEQ ID NO372 GPPGEK SEQ ID NO640 GPPGLM SEQ ID NO643 PGPMGP SEQ ID NO25 .LPGPDG SEQ ID NO2214 .LRGIPG SEQ ID NO760 .PGFPGA SEQ ID NO2218 .QQGNRG SEQ ID NO2221 .VGQPGP SEQ ID NO2222 .GEPGMQ SEQ ID NO2224 DGLPGS SEQ ID NO611 PGPPGL SEQ ID NO614 GLPGQQ SEQ ID NO616 PGLPGI SEQ ID NO504 PGMQGE SEQ ID NO620 RGPPGP SEQ ID NO148 GPLGEK SEQ ID NO625 LQGIRG SEQ ID NO628 PGLPGP SEQ ID NO58 GAPGPQ SEQ ID NO445 DRGPQG SEQ ID NO442 EKGQKG SEQ ID NO637 DGGVPN SEQ ID NO638 AEGLPG SEQ ID NO641 LPGFAG SEQ ID NO644 .PGIPGT SEQ ID NO2227 .PGDIVF SEQ ID NO2215 .GNKGDP SEQ ID NO2216 .PGPRGK SEQ ID NO2219 .PGPPGP SEQ ID NO458 .SKGEKG SEQ ID NO2226 .GEPGPP SEQ ID NO675 or with any of the following sequences at the C-terminal of a peptide:

TABLE 16 C-terminal sequences of protease generated peptide fragments of Collagen type IV. Collagen type IV RGPPGP SEQ ID NO148 VDHGFL SEQ ID NO648 PGLPGS SEQ ID NO651 LPGSMG SEQ ID NO654 PGLPGE SEQ ID NO656 GPPGLV SEQ ID NO658 PGLPGP SEQ ID NO58 PGPRGE SEQ ID NO663 GIGPPG SEQ ID NO611 PGAIGP SEQ ID NO668 GPKGPP SEQ ID NO671 GSVGYP SEQ ID NO673 GEPGPP SEQ ID NO675 PGYTNG SEQ ID NO678 LSGPPG SEQ ID NO680 GVSGPK SEQ ID NO682 HVPGML. SEQ ID NO2228 GVPGQA. SEQ ID NO2231 QEGPLG. SEQ ID NO2234 NRGLGF. SEQ ID NO2238 GEKGNK. SEQ ID NO2241 PPGPKG. SEQ ID NO167 GEKGSK. SEQ ID NO2246 AGIPQK. SEQ ID NO2237 SVDHGF SEQ ID NO646 PGQPGY SEQ ID NO649 GTPSVD SEQ ID NO652 GPPGVP SEQ ID NO633 GDPGPP SEQ ID NO373 PGIPGP SEQ ID NO659 DGIPGP SEQ ID NO661 GQGPPG SEQ ID NO664 PGIDGV SEQ ID NO666 PPGPPG SEQ ID NO119 SPGPPG SEQ ID NO672 PQGPPG SEQ ID NO389 AGNPGP SEQ ID NO676 FPGPQG SEQ ID NO679 PGAPGL SEQ ID NO467 PGPPGP SEQ ID NO458 LPVPGQ. SEQ ID NO2229 VPGQAQ. SEQ ID NO2232 LPGEVL. SEQ ID NO2235 IPSDTL. SEQ ID NO2239 PVGPPG. SEQ ID NO2242 RGKPGM. SEQ ID NO2244 EKGSKG. SEQ ID NO2247 PSVDHG SEQ ID NO647 QPGYTN SEQ ID NO650 SVGSPG SEQ ID NO653 PGFPGL SEQ ID NO655 HQGEMG SEQ ID NO657 PGFPGT SEQ ID NO660 SGPKGY SEQ ID NO662 KVDMGS SEQ ID NO665 KGHMGE SEQ ID NO667 KGLPGP SEQ ID NO669 GPKGLP SEQ ID NO670 PPGSPG SEQ ID NO674 PGIKGS SEQ ID NO677 PGPQGP SEQ ID NO453 GVMGTP SEQ ID NO681 LGPPGL. SEQ ID NO2230 GPDGFL. SEQ ID NO2233 RGIPGF. SEQ ID NO2236 PAGEKG. SEQ ID NO2240 DIVFRK. SEQ ID NO2243 PGTRGL. SEQ ID NO2245 SGQPGL. SEQ ID NO2248

Collagen V

We have determined that the enzymes listed in the following table cleave type v collagen at least the following cleavage sites (marked “.” or in the absence of a ‘.’, at the end of the sequence):

TABLE 14A Cleavage fragments of collagen type V Protease Neo-epitope (COV) MMP2, Alpha3 K.GDPGPPGPIGSLG.H SEQ ID NO683 MMP2, Alpha3 G.LRGIPGPVGEPG.L SEQ ID NO684 MMP2, Alpha3 V.IGPPGLQGLPGPPGE.K SEQ ID NO685 MMP2, Alpha3 G.KDGIPGPLGPLGPPG.A SEQ ID NO686 MMP2, Alpha3 G.LRGIPGPVGEPGLL.G SEQ ID NO687 MMP2, Alpha3 G.VLGPQGKTGEVGPLG.E SEQ ID NO688 MMP2, Alpha3 K.DGIPGPLGPLGPPGAA.G SEQ ID NO689 MMP2, Alpha3 G.EDGERGAEGPPGPTG.Q SEQ ID NO690 MMP2, Alpha3 G.LQGPPGFPGPKGPPG.H SEQ ID NO691 MMP2, Alpha3 P.IGSLGHPGPPGVAGPLG.Q SEQ ID NO692 MMP2, Alpha3 G.IRGPPGTVIMMPFQ.F SEQ ID NO693 MMP2, Alpha3 G.QMGPPGPLGPSGLPGLK.G SEQ ID NO694 MMP2, Alpha3 G.LLGAPGQMGPPGPLGPSG.L SEQ ID NO695 MMP2, Alpha3 G.LRGIPGPVGEPGLLGAPG.Q SEQ ID NO696 MMP2, Alpha3 G.LLGPRGSPGPTGRPGVTG.I SEQ ID NO697 MMP2, Alpha3 G.IRGPPGTVIMMPFQF.A SEQ ID NO698 MMP2, Alpha3 G.KDGIPGPLGPLGPPGAAGP.S SEQ ID NO699 MMP2, Alpha3 G.KDGIPGPLGPLGPPGAAGPSG.E SEQ ID NO700 MMP2, Alpha3 Q.GLPGLEGREGAKGELGPPGPLG.K SEQ ID NO701 MMP2, Alpha3 L.GPIGEKGKSGKTGQPGLEGERGPPGSRG.E SEQ ID NO702 MMP2, Alpha3 G.LRGIPGPVGEPGLLGAPGQMGPPGPLGPSG.L SEQ ID NO703 MMP2, Alpha3 G.ANGSPGERGPLGPAGGIGLPGQSGSEGPVGPAG.K SEQ ID NO704 MMP2, Alpha3 G.LIGTPGEKGPPGNPGIPGLPGSDGPLGHPGHEGPTG.E SEQ ID NO705 MMP2, Alpha1 G.LPGEPGPRG.L SEQ ID NO706 MMP2, Alpha1 L.ALRGPAGPMG.L SEQ ID NO707 MMP2, Alpha1 R.LALRGPAGPMG.L SEQ ID NO708 MMP2, Alpha1 G.LTGRPGPVGPPGSGG.L SEQ ID NO709 MMP2, Alpha1 G.LLGPKGPPGPPGPPG.V SEQ ID NO710 MMP2, Alpha1 G.IPGRPGPQGPPGPAG.E SEQ ID NO711 MMP2, Alpha1 P.GPDGPPGPMGPPGLP.G SEQ ID NO712 MMP2, Alpha1 G.QPGPSGADGEPGPRG.Q SEQ ID NO713 MMP2, Alpha1 G.ETGFQGKTGPPGPPG.V SEQ ID NO714 MMP2, Alpha1 G.LRGFPGDRGLPGPV.G SEQ ID NO715 MMP2, Alpha1 G.LRGFPGDRGLPGPVG.A SEQ ID NO716 MMP2, Alpha1 G.KTGPIGPQGAPGKPGPDG.L SEQ ID NO717 MMP2, Alpha1 G.PPGRPGLPGADGLPGPPG.T SEQ ID NO718 MMP2, Alpha1 G.LKGNEGPPGPPGPAGSPGE.R SEQ ID NO719 MMP2, Alpha1 G.LRGFPGDRGLPGPVGALG.L SEQ ID NO720 MMP2, Alpha1 G.ERGHPGPPGPPGEQGLPG.L SEQ ID NO721 MMP2, Alpha1 I.GPPGEQGEKGDRGLPGPQG.S SEQ ID NO722 MMP2, Alpha1 G.EAGHPGPPGPPGPPGEVIQPLP.I SEQ ID NO723 MMP2, Alpha1 K.PGPKGNSGGDGPAGPPGERGPNGP.Q SEQ ID NO724 MMP2, Alpha1 G.EQGLPGSPGPDGPPGPMGPPGLPG.L SEQ ID NO725 MMP2, Alpha1 E.GPPGEKGGQGPPGPQGPIGYPGPRG.V SEQ ID NO726 MMP2, Alpha1 G.FPGPKGPPGPPGKDGLPGHPGQRG.E SEQ ID NO727 MMP2 L.PFRFGGGGDA SEQ ID NO728 MMP2 and 9 GSKGPMVSAQ.E SEQ ID NO729 MMP2 and 9 Q.ESQAQAILQQ SEQ ID NO730 MMP9, Alpha1 L.ALRGPAGPMG.L SEQ ID NO707 MMP9, Alpha1 G.AIGPPGEKGPLG.K SEQ ID NO731 MMP9, Alpha1 G.GPNGDPGPLGPPG.E SEQ ID NO732 MMP9, Alpha1 P.PGPPGEQGLPGL.A SEQ ID NO733 MMP9, Alpha1 G.LLGPKGPPGPPGPPG.V SEQ ID NO734 MMP9, Alpha1 G.IPGRPGPQGPPGPAG.E SEQ ID NO711 MMP9, Alpha1 G.QPGPSGADGEPGPRG.Q SEQ ID NO713 MMP9, Alpha1 G.QQGNPGAQGLPGPQG.A SEQ ID NO735 MMP9, Alpha1 G.KEGPPGEKGGQGPPG.P SEQ ID NO736 MMP9, Alpha1 G.ETGFQGKTGPPGPPG.V SEQ ID NO737 MMP9, Alpha1 G.EKGHPGLIGLIGPPG.E SEQ ID NO738 MMP9, Alpha1 G.LRGFPGDRGLPGPVG.A SEQ ID NO716 MMP9, Alpha1 G.KTGPIGPQGAPGKPGPDG.L SEQ ID NO739 MMP9, Alpha1 P.GPDGPPGPMGPPGLPGLK.G SEQ ID NO740 MMP9, Alpha1 G.ERGHPGPPGPPGEQGLPG.L SEQ ID NO721 MMP9, Alpha1 G.ERGPNGPQGPTGFPGPKGPPGPPG.K SEQ ID NO741 MMP9, Alpha1 L.IGLIGPPGEQGEKGDRGLPGPQGS.S SEQ ID NO742 MMP9, Alpha1 E.GPPGEKGGQGPPGPQGPIGYPGPRG.V SEQ ID NO726 MMP9, Alpha1 I.GPPGPPGLPGPPGPKGAKGSSGPTGPKGE.A SEQ ID NO743 MMP9, Alpha1 P.LGPPGEKGKLGVPGLPGYPGRQGPKGSI.G SEQ ID NO744 MMP9, Alpha1 Q.GPKGSIGFPGFPGANGEKGGRGTPGKPGPRG.Q SEQ ID NO745 MMP9, Alpha3 P.GPKGDPGPPGPIG.S SEQ ID NO746 MMP9, Alpha3 K.GDPGPPGPIGSLG.H SEQ ID NO683 MMP9, Alpha3 A.PGIPGEKGLPGL.Q SEQ ID NO747 MMP9, Alpha3 Q.GPPGPKGDPGPPGP.I SEQ ID NO748 MMP9, Alpha3 G.SLGHPGPPGVAGPLG.Q SEQ ID NO749 MMP9, Alpha3 G.KDGIPGPLGPLGPPG.A SEQ ID NO686 MMP9, Alpha3 G.VLGPQGKTGEVGPLG.E SEQ ID NO688 MMP9, Alpha3 G.ELGFQGQTGPPGPAG.V SEQ ID NO750 MMP9, Alpha3 G.EDGERGAEGPPGPTG.Q SEQ ID NO690 MMP9, Alpha3 G.LQGPPGFPGPKGPPG.H SEQ ID NO691 MMP9, Alpha3 G.EKGHIGLIGLIGPPG.E SEQ ID NO751 MMP9, Alpha3 G.QMGPPGPLGPSGLPGLK.G) SEQ ID NO694 MMP9, Alpha3 G.PVGEPGLLGAPGQMGPPG.P SEQ ID NO752 MMP9, Alpha3 G.LRGIPGPVGEPGLLGAPG.Q SEQ ID NO696 MMP9, Alpha3 G.LLGPRGSPGPTGRPGVTG.I SEQ ID NO697 MMP9, Alpha3 G.KDGIPGPLGPLGPPGAAGPSG.E SEQ ID NO700 MMP9, Alpha3 Q.GLPGLEGREGAKGELGPPGPLG.K SEQ ID NO701 MMP9, Alpha3 G.SRGERGPPGPTGKDGIPGPLGPLG.P SEQ ID NO753 MMP9, Alpha3 G.EKGKSGKTGQPGLEGERGPPGSRG.E SEQ ID NO754 MMP9, Alpha3 L.GPIGEKGKSGKTGQPGLEGERGPPGSRG.E SEQ ID NO702 MMP9, Alpha3 G.ANGSPGERGPLGPAGGIGLPGQSGSEGPVGPAG.K SEQ ID NO704 MMP9, Alpha3 G.LIGTPGEKGPPGNPGIPGLPGSDGPLGHPGHEGPTG.E SEQ ID NO705 MMP13, Alpha1 L.PGEPGPRG.L SEQ ID NO755 MMP13, Alpha1 A.LRGPAGPMG.L SEQ ID NO756 MMP13, Alpha1 G.LPGEPGPRG.L SEQ ID NO706 MMP13, Alpha1 L.ALRGPAGPMG.L SEQ ID NO707 MMP13, Alpha1 R.LALRGPAGPMG.L SEQ ID NO708 MMP13, Alpha1 G.LRGFPGDRGLPGPVG.A SEQ ID NO716 MMP13, Alpha1 Q.ESQAQAILQQARLA.L SEQ ID NO730 MMP13, Alpha1 P.GPDGPPGPMGPPGLPGLK.G SEQ ID NO740 MMP13, Alpha1 G.PQGAIGPPGEKGPLGKPGLPGMPGADGPPGHPG.K SEQ ID NO757 MMP13, Alpha1 A.GPMGLTGRPGPVGPPGSGGLKGEPGDVGPQGPRG.V SEQ ID NO758 MMP13, Alpha3 G.VLGPQGKTGEVGPLG.E SEQ ID NO688 MMP13, Alpha3 G.LRGIPGPVGEPGLLGAPG.Q SEQ ID NO696 MMP13, Alpha3 G.LRGIPGPVGEPGLLGAPGQMGPPGPLGPSG.L SEQ ID NO703 MMP13, Alpha3 G.LRGIPGPVGEPGLLGAPGQMGPPGPLGPSGLPG.L SEQ ID NO759 P is hydroxyproline, K indicates hydroxylysine, glycosylation, lipoxidation or cross linking.

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type v collagen.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 15a N-terminal sequences of protease generated peptide fragments of Collagen type V. Collagen type V GDPGPP SEQ ID NO373 LQGPPG SEQ ID NO62 ANGSPG SEQ ID NO764 IPGRPG SEQ ID NO767 LKGNEG SEQ ID NO770 FPGPKG SEQ ID NO491 LLGPKG SEQ ID NO775 IGLIGP SEQ ID NO778 GPPGPK SEQ ID NO780 PVGEPG SEQ ID NO783 ANGSPG SEQ ID NO764 LLGAPG SEQ ID NO785 LALRGP SEQ ID NO788 LRGFPG SEQ ID NO790 PGPKGN SEQ ID NO527 AIGGPP SEQ ID NO794 LLGPRG SEQ ID NO797 GPKGDP SEQ ID NO799 LQGPPG SEQ ID NO62 SRGERG SEQ ID NO803 LPGEPG SEQ ID NO766 QMGPPG SEQ ID NO802 ALRGPA SEQ ID NO810 KDGIP SEQ ID NO813 GPKGSI SEQ ID NO816 LRGIPG SEQ ID NO760 IGSLGH SEQ ID NO762 LIGTPG SEQ ID NO765 GPDGPP SEQ ID NO768 ERGHPG SEQ ID NO771 PFRFGG SEQ ID NO773 QQGNPG SEQ ID NO776 GPPGPP SEQ ID NO100 SLGHPG SEQ ID NO781 LRGIPG SEQ ID NO760 LIGTPG SEQ ID NO765 GLPGLE SEQ ID NO786 LTGRPG SEQ ID NO789 KTGPIG SEQ ID NO791 EQGLPG SEQ ID NO793 GPNGDP SEQ ID NO795 GPDGPP SEQ ID NO768 GDPGPP SEQ ID NO373 EKGHIG SEQ ID NO801 EKGKSG SEQ ID NO804 PQGAIG SEQ ID NO806 ETGFQG SEQ ID NO808 EAGHPG SEQ ID NO811 VLGPQG SEQ ID NO814 ELGFQG SEQ ID NO817 IGPPGI SEQ ID NO761 IRGPPG SEQ ID NO763 LPGEPG SEQ ID NO766 QPGPSG SEQ ID NO769 GPPGEQ SEQ ID NO772 ESQAQA SEQ ID NO774 KEGPPG SEQ ID NO777 LGPPGE SEQ ID NO779 KDGIPG SEQ ID NO782 KDGIPG SEQ ID NO782 PGEPGP SEQ ID NO784 GIPGEK SEQ ID NO787 LLGPKG SEQ ID NO775 PPGRPG SEQ ID NO792 GPPGEK SEQ ID NO640 PGPPGE SEQ ID NO796 ERGPNG SEQ ID NO798 PGIPGE SEQ ID NO800 QMGPPG SEQ ID NO802 GPIGEK SEQ ID NO805 GPMGLT SEQ ID NO807 GSKGPM SEQ ID NO809 EKGHPG SEQ ID NO812 EDGERG SEQ ID NO815 LRGPAG SEQ ID NO818 P is hydroxyproline, K indicates hydroxylysine, glycosylation, lipoxidation or cross linking. or with any of the following sequences at the C-terminal of a peptide:

TABLE 16a C-terminal sequences of protease generated peptide fragments of Collagen type V. Collagen type V PIGSLG SEQ ID NO819 PPGPTG SEQ ID NO820 RPGVTG SEQ ID NO823 PVGPAG SEQ ID NO826 GPPGLP SEQ ID NO828 AGSPGE SEQ ID NO829 YPGPRG SEQ ID NO831 LIGPPG SEQ ID NO834 PPGPIG SEQ ID NO174 QMGPPG SEQ ID NO802 QQARLA SEQ ID NO837 PLGPPG SEQ ID NO839 IMMPFQ SEQ ID NO842 AAGPSG SEQ ID NO844 PAGPMG SEQ ID NO847 GLPGPV SEQ ID NO849 LPGPQG SEQ ID NO852 PGPQGS SEQ ID NO855 GPPGAA SEQ ID NO857 PPGPAG SEQ ID NO52 QGLPGL SEQ ID NO859 PPGSRG SEQ ID NO846 PPGPLG SEQ ID NO845 PLGPLG SEQ ID NO863 PVGEPG SEQ ID NO783 PKGPPG SEQ ID NO821 MMPFQF SEQ ID NO824 HEGPTG SEQ ID NO827 GQGPPG SEQ ID NO664 PVGALG SEQ ID NO830 HPGQRG SEQ ID NO832 GLPGLK SEQ ID NO835 KGLPGL SEQ ID NO836 LLGAPG SEQ ID NO785 PPGHPG SEQ ID NO838 GEPGLL SEQ ID NO840 GLPGLK SEQ ID NO835 PPGPLG SEQ ID NO845 PPGSGG SEQ ID NO848 LPGPVG SEQ ID NO850 VIQPLP SEQ ID NO853 EKGPLG SEQ ID NO856 TGPKGE SEQ ID NO858 PPGPAG SEQ ID NO52 PSGLPG SEQ ID NO860 LPGPPG SEQ ID NO72 KPGPRG SEQ ID NO862 PPGSRG SEQ ID NO846 PGPPGE SEQ ID NO796 VAGPLG SEQ ID NO822 PGAAGP SEQ ID NO825 EPGPRG SEQ ID NO516 PPGPPG SEQ ID NO119 EQGLPG SEQ ID NO793 GGGGDA SEQ ID NO833 PPGPPG SEQ ID NO119 PGPPGP SEQ ID NO458 RPGVTG SEQ ID NO823 PQGPRG SEQ ID NO150 EVGPLG SEQ ID NO841 PLGPSG SEQ ID NO843 PPGSRG SEQ ID NO846 PPGPPG SEQ ID NO119 KPGPDG SEQ ID NO851 RGPNGP SEQ ID NO854 PLGPPG SEQ ID NO839 GPKGSI SEQ ID NO816 PPGPTG SEQ ID NO820 LLGAPG SEQ ID NO785 PPGLPG SEQ ID NO861 PKGPPG SEQ ID NO821 P is hydroxyproline, K indicates hydroxylysine, glycosylation, lipoxidation or cross linking.

Collagen VI

We have determined that the enzymes listed in the following table cleave type vi collagen at least the following cleavage sites (marked “.” or in the absence of a ‘.’, at the end of the sequence):

TABLE 14B Cleavage fragments of collagen type VI Protease Neoepitope MMP2 G.YRGPEGPQGPPG.H SEQ ID NO864 MMP2 G.PIGPKGYRGDEGPP.G SEQ ID NO865 MMP2, I.GIGIGNADIT.E SEQ ID NO866 (a3) MMP2, G.AQGPAGPAGPPG.L SEQ ID NO867 (a3) MMP9 G.LIGEQGISGPRG.S SEQ ID NO868 MMP9 P.PGLIGEQGISGPR.G SEQ ID NO869 MMP9 E.PGEPGPKGGIGNRG.P SEQ ID NO870 MMP9 G.ISGPRGSGGAAGAPGERGRTGPLG.R SEQ ID NO871 MMP13 PGPAGPPGDPGLMG SEQ ID NO872 FAP-1 VAAKPAAVRPPAAAAAKPVATKPEVPRP SEQ ID NO873 FAP-1 GEPGLNGTTGPKGI SEQ ID NO874 FAP-1 IGPKGIPGEDGYRGYPG SEQ ID NO875 FAP-1 VAVVQHAPSESVDNASMPPVKVEFSL SEQ ID NO876 FAP-2 LGPMGVPGRD SEQ ID NO877 FAP-2 GEPGPPGEKGEAGDEGNPGPDGAPGERG SEQ ID NO878 FAP-2 RGPIGSIGPKGIPGEDGYRGYPGDEGGP SEQ ID NO879 FAP-2 PPPPQPARSAS SEQ ID NO880 FAP-2 FGPSAATPAPPG SEQ ID NO881 FAP-2 GPKGETGDLGPMGVPGRDGVPGGPGETGK SEQ ID NO882

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type v collagen.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 15b N-terminal sequences of protease generated peptide fragments of Collagen type VI. Collagen type VI YRGPEG SEQ ID NO883 ISGPRG SEQ ID NO886 GEPGPP SEQ ID NO675 AQGPAG SEQ ID NO891 GEPGLN SEQ ID NO894 FGPSAA SEQ ID NO897 LGPMGV SEQ ID NO899 PIGPKG SEQ ID NO865 PGPAGP SEQ ID NO887 RGPIGS SEQ ID NO889 LIGEQG SEQ ID NO892 IGPKGI SEQ ID NO895 GPKGET SEQ ID NO898 GIGIGN SEQ ID NO885 VAAKPA SEQ ID NO888 PPPPQP SEQ ID NO890 PGLIGE SEQ ID NO893 VAVVQH SEQ ID NO896 PGEPGP SEQ ID NO784 or with any of the following sequences at the C-terminal of a peptide:

TABLE 16b C-terminal sequences of protease generated peptide fragments of Collagen type VI. Collagen type VI GDEGPP SEQ ID NO900 DPGLMG SEQ ID NO902 GDEGGP SEQ ID NO905 ISGPRG SEQ ID NO886 YRGYPG SEQ ID NO909 PGETGK SEQ ID NO912 GNADIT SEQ ID NO901 PEVPRP SEQ ID NO903 PARSAS SEQ ID NO906 GISGPR SEQ ID NO907 KVEFSL SEQ ID NO910 RTGPLG SEQ ID NO913 PAGPPG SEQ ID NO133 TGPKGI SEQ ID NO904 TPAPPG SEQ ID NO915 GIGNRG SEQ ID NO908 GVPGRD SEQ ID NO911 APGERG SEQ ID NO914

Proteoglycans

In another aspect of the invention, said peptide fragments are fragments of proteoglycans versican, lumican, perlecan, biglycan and decorin, which are all identified in fibrotic tissue.

Several candidate proteases may be responsible for the digestion of proteoglycans in fibrotic lesions We have determined that the enzymes listed in table 17 generate lumican, versican, biglycan, perlecan and decorin resulting in at least following cleavage products:

TABLE 17 Cleavage fragments of biglycan, decorin, versican, lumican, and perlecan. Protease Biglycan MMP-3 SVPKEISPDTTLLDLQNNDISE SEQ ID NO916 MMP-3 KSVPKEISPDTTLLDLQNNDISE SEQ ID NO917 MMP-9 NSGFEPGAFDGLKLNYLRISEAK SEQ ID NO918 MMP-9 LKSVPKEISPDTTLLDLQNNDISE SEQ ID NO919 MMP-12 LRISEAKLTGIPKDLPET SEQ ID NO920 MMP-13 LKSVPKEISPDTTLLDLQNNDISE SEQ ID NO919 MMP-13 LTGIPKDLPETLNELHLDHNKIQAIE SEQ ID NO921 ADAMTS4 RISEAKLTGIPKDLPETLNE SEQ ID NO922 ADAMTS4 AIELEDLLRYSK SEQ ID NO923 ADAMTS4 AIELEDLLRY SEQ ID NO924 ADAMTS4 EAKLTGIPKDLPETLNE SEQ ID NO925 ADAMTS4 LKAVPKEISPDTTLLDLQNNDISE SEQ ID NO926 MMP-8 LLDLQNNDISELRKDD SEQ ID NO927 MMP-8 IELEDLLRYS SEQ ID NO928 CathepsinS NSGFEPGAFDGLK SEQ ID NO929 Protease Decorin MMP-12 IVIELGTNPLK SEQ ID NO930 MMP-3 DEASGIGPEVPDDR SEQ ID NO931 MMP-3 LHLDGNKISRVDAAS SEQ ID NO932 MMP-3 VNNKISKVSPGAFTPL SEQ ID NO933 MMP-3 LILVNNKISKVSPGAFTPLVKLER SEQ ID NO934 MMP-9 SNPVQYWEIQPSTFR SEQ ID NO935 CathepsinK SSGIENGAFQGMK SEQ ID NO884 CathepsinK SSGIENGAFQGMKKLS SEQ ID NO946 ADAMTS1 KITEIKDGDFK SEQ ID NO936 ADAMTS1 GLPPSLTELHLDGNK SEQ ID NO937 Versican Unknown LLASDAGLYR SEQ ID NO938 Unknown LATVGELQAAWR SEQ ID NO939 Unknown ETTVLVAQNGNIK SEQ ID NO940 Lumican Unknown SLEDLQLTHNK SEQ ID NO941 Unknown LKEDAVSAAFK SEQ ID NO942 Perlecan Unknown SIEYSPQLEDAGSR SEQ ID NO943 Unknown LEGDTLIIPR SEQ ID NO944 ADAMTS4 VSEAVVEKLEPEYR SEQ ID NO945 ADAMTS4 EVSEAVVEKLEPEYR SEQ ID NO947 ADAMTS4 SIEYSPQLEDASAKEFR SEQ ID NO948

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of type versican, lumican, decorin, perlecan, and biglycan.

Suitable immunological binding partners may therefore be specifically reactive with any of the following at the N terminal of a peptide:

TABLE 18 N-terminal sequences of protease generated peptide fragments of biglycan, decorin, lumican, versican, and perlecan. Biglycan SVPKEI SEQ ID NO949 NSGFEP SEQ ID NO952 IELEDL SEQ ID NO957 LRISEA SEQ ID NO958 LLDLQN SEQ ID NO961 GLKLNY SEQ ID NO950 LKSVPK SEQ ID NO953 QCSDLG SEQ ID NO955 LTGIPK SEQ ID NO959 RISEAK SEQ ID NO951 AIELED SEQ ID NO954 EAKLTG SEQ ID NO956 LKAVPK SEQ ID NO960 Decorin IVIELG SEQ ID NO962 NGLNQM SEQ ID NO965 SSGIEN SEQ ID NO968 SNPVQY SEQ ID NO971 DEASGI SEQ ID NO963 LHLDGN SEQ ID NO966 KITEIK SEQ ID NO969 VNNKIS SEQ ID NO964 LILVNN SEQ ID NO967 GLPPSL SEQ ID NO970 Versican LLASDA SEQ ID NO972 ENQDAR SEQ ID NO975 LATVGE SEQ ID NO973 NGFDQC SEQ ID NO976 ETTVLV SEQ ID NO974 SLTVVK SEQ ID NO977 Lumican SLEDLQ SEQ ID NO978 LQHNRL SEQ ID NO985 LKEDAV SEQ ID NO979 HLQHNR SEQ ID NO980 Perlecan SIEYSP SEQ ID NO981 EVSEAV SEQ ID NO984 LVNFTR SEQ ID NO982 VSEAVV SEQ ID NO983 or with any of the following sequences in table 19, at the C-terminal of a peptide:

TABLE 19 C-terminal sequences of protease generated peptide fragments of biglycan, decorin, lumican, versican, and perlecan. Biglycan NNDISE SEQ ID NO986 RISEAK SEQ ID NO951 LRKDDF SEQ ID NO991 KDLPET SEQ ID NO994 LNELHL SEQ ID NO997 YWEVQP SEQ ID NO987 KIQAIE SEQ ID NO989 LLRYSK SEQ ID NO992 DLLRYS SEQ ID NO995 EDLLRY SEQ ID NO988 PETLNE SEQ ID NO990 ELRKDD SEQ ID NO993 AFDGLK SEQ ID NO996 Decorin GTNPLK SEQ ID NO998 SSGIEN SEQ ID NO968 GMKKLS SEQ ID NO1003 QPSTFR SEQ ID NO1006 EVPDDR SEQ ID NO999 RVDAAS SEQ ID NO1001 KDGDFK SEQ ID NO1004 AFQGMK SEQ ID NO1007 GAFTPL SEQ ID NO1000 LVKLER SEQ ID NO1002 HLDGNK SEQ ID NO1005 Versican CDVMYG SEQ ID NO1008 IGQDYK SEQ ID NO1010 NGFDQC SEQ ID NO976 QNGINK SEQ ID NO1009 Lumican QLTHNK SEQ ID NO1011 VSAAFK SEQ ID NO1012 GLKSLE SEQ ID NO1013 Perlecan EDAGSR SEQ ID NO1014 SAKEFR SEQ ID NO1017 EFREVS SEQ ID NO1015 LEPEYR SEQ ID NO1018 VAQQDS SEQ ID NO1016

CRP

Several candidate proteases may be responsible for the digestion of CRP in fibrotic tissue the literature reports many different proteases in fibrotic tissue. Most likely, this is the result of the large range of complicated processes eventually leading to fibrosis. However, in our assessment, early phases may consist of a range of MMPs, whereas later stages may rely more on cathepsin K degradation of the matrix, resulting in different neo-epitope profiles dependent on the levels of disease. We have through a range of in vitro cleavages of pure native proteins determined that the enzymes listed in the following tables of cleaved CRP at least following cleavage sites (marked * in Table 20, but at the ends of each sequence in Table 21):

TABLE 20 CRP fragments generated by specific proteases. Protease/Protein Neo-epitope CRP + CatK K*ESDTSYVSLKAPLT*K SEQ ID NO1019 CRP + CatK G*GNFEGSQSLVGDIG*N SEQ ID NO1020 CRP + MMP9 A*LKYEVQGEVFTKPQ*L SEQ ID NO1021 CRP + MMP9 G*IVEFWVDGKPRV*R SEQ ID NO1022 CRP + MMP1/MMP3 R*KAFVFPKE*S SEQ ID NO1023 CRP + MMP3 K*YEVQGEVFTKPQLWP*- SEQ ID NO1024 CRP + MMP3 D*SFGGNFEGSQS*L SEQ ID NO1025 CRP + MMP3 D*FVLSPDEINT*I SEQ ID NO1026 CRP + MMP3 S*LKKGYTVGAEA*S SEQ ID NO1027 CRP + MMP3 A*FGQTDMSRKA*F SEQ ID NO1028 CRP + MMP3 S*LKKGYTVGAEAS*I SEQ ID NO1029 CRP + MMP3 G*EVFTKPQLWP*- SEQ ID NO1030 CRP + MMP3 S*IILGQEQDSFGGNF SEQ ID NO1031 CRP + MMP3 K*YEVQGEVFTKPQ.L SEQ ID NO1032

TABLE 21 CRP fragments generated by specific proteases. Protease Neoepitope Aminoacid Nos* MMP9 AFVFPK SEQ ID NO1033 026-031 MMP9 FGQTDMSR SEQ ID NO1034 017-024 MMP9 FGQTDMSRK SEQ ID NO1035 017-025 MMP9 FGQTDMSRKA SEQ ID NO1036 017-026 MMP9 FGQTDMSRKAF SEQ ID NO1037 017-027 MMP9 FGQTDMSRKAFVFPKE SEQ ID NO1038 017-032 MMP9 FGQTDMSRKAFVFPKESDTS SEQ ID NO1039 017-036 MMP9 FGQTDMSRKAFVFPKESDTSYV SEQ ID NO1040 017-038 MMP9 FGQTDMSRKAFVFPKESDTSYVS SEQ ID NO1041 017-039 MMP9 TDMSRKAFVFPKESDTSYV SEQ ID NO1042 020-038 MMP9 MSRKAFVFPKESDTS SEQ ID NO1043 022-036 MMP9 SRKAFVFPKESDTSYV SEQ ID NO1044 023-038 MMP9 RKAFVFPKE SEQ ID NO1045 024-032 MMP9 RKAFVFPKESDTSYV SEQ ID NO1046 024-038 MMP9 RKAFVFPKESDTSYVS SEQ ID NO1047 024-039 MMP9 KAFVFPKE SEQ ID NO1048 025-032 MMP9 KAFVFPKESD SEQ ID NO1049 025-034 MMP9 KAFVFPKESDT SEQ ID NO1050 025-035 MMP9 KAFVFPKESDTS SEQ ID NO1051 025-036 MMP9 KAFVFPKESDTSYV SEQ ID NO1052 025-038 MMP9 KAFVFPKESDTSYVS SEQ ID NO1053 025-039 MMP9 AFVFPKE SEQ ID NO1054 026-032 MMP9 AFVFPKESDT SEQ ID NO1055 026-035 MMP9 AFVFPKESDTSYV SEQ ID NO1056 026-038 MMP9 AFVFPKESDTSYVS SEQ ID NO1057 026-039 MMP9 AFVFPKESDTSYVSL SEQ ID NO1058 026-040 MMP9 FVFPK SEQ ID NO1059 027-031 MMP9 FVFPKE SEQ ID NO1060 027-032 MMP9 FVFPKESD SEQ ID NO1061 027-034 MMP9 FVFPKESDTS SEQ ID NO1062 027-036 MMP9 FVFPKESDTSY SEQ ID NO1063 027-037 MMP9 FVFPKESDTSYV SEQ ID NO1064 027-038 MMP9 FVFPKESDTSYVS SEQ ID NO1065 027-039 MMP9 FVFPKESDTSYVSL SEQ ID NO1066 027-040 MMP9 VFPKESDTS SEQ ID NO1067 028-036 MMP9 VFPKESDTSYV SEQ ID NO1068 028-038 MMP9 VFPKESDTSYVS SEQ ID NO1069 028-039 MMP9 VFPKESDTSYVSL SEQ ID NO1070 028-040 MMP9 FPKESDTSYVS SEQ ID NO1071 029-039 MMP9 KESDTSYVSLKAPLTKP SEQ ID NO1072 031-047 MMP9 SDTSYVSLKAPLTKP SEQ ID NO1073 033-047 MMP9 SLKAPLTKP SEQ ID NO1074 039-047 MMP9 SLKAPLTKPLK SEQ ID NO1075 039-049 MMP9 LKAPLTKPLK SEQ ID NO1076 040-049 MMP9 FYTELSSTRGYS SEQ ID NO1077 057-068 MMP9 LSSTRGYS SEQ ID NO1078 061-068 MMP9 SSTRGYS SEQ ID NO1079 062-068 MMP9 STRGYS SEQ ID NO1080 063-068 MMP9 IFSYATKRQ SEQ ID NO1081 069-077 MMP9 IFSYATKRQDNEILI SEQ ID NO1082 069-083 MMP9 SYATKRQDNEILI SEQ ID NO1083 071-083 MMP9 YATKRQDNEIL SEQ ID NO1084 072-082 MMP9 YATKRQDNEILI SEQ ID NO1085 072-083 MMP9 YATKRQDNEILIF SEQ ID NO1086 072-084 MMP9 TKRQDNEILI SEQ ID NO1087 074-083 MMP9 TKRQDNEILIF SEQ ID NO1088 074-084 MMP9 TKRQDNEILIFWSKDI SEQ ID NO1089 074-089 MMP9 KRQDNEILI SEQ ID NO1090 075-083 MMP9 KRQDNEILIF SEQ ID NO1091 075-084 MMP9 WSKDIGYS SEQ ID NO1092 085-092 MMP9 SKDIGYS SEQ ID NO1093 086-092 MMP9 IVEFWVDGKPRV SEQ ID NO1094 124-135 MMP9 EFWVDGKPR SEQ ID NO1095 126-134 MMP9 WVDGKPRV SEQ ID NO1096 128-135 MMP9 VDGKPRV SEQ ID NO1097 129-135 MMP9 SLKKGYTVGAE SEQ ID NO1098 138-148 MMP9 SLKKGYTVGAEA SEQ ID NO1099 138-149 MMP9 SLKKGYTVGAEAS SEQ ID NO1100 138-150 MMP9 LKKGYTV SEQ ID NO1101 139-145 MMP9 LKKGYTVG SEQ ID NO1102 139-146 MMP9 LKKGYTVGA SEQ ID NO1103 139-147 MMP9 LKKGYTVGAE SEQ ID NO1104 139-148 MMP9 LKKGYTVGAEA SEQ ID NO1105 139-149 MMP9 LKKGYTVGAEAS SEQ ID NO1106 139-150 MMP9 LKKGYTVGAEASI SEQ ID NO1107 139-151 MMP9 SIILGQEQDSFGGN SEQ ID NO1108 150-163 MMP9 SIILGQEQDSFGGNFEGSQ SEQ ID NO1109 150-168 MMP9 SIILGQEQDSFGGNFEGSQS SEQ ID NO1110 150-169 MMP9 IILGQEQDSFGGNFEGS SEQ ID NO1111 151-067 MMP9 IILGQEQDSFGGNFEGSQS SEQ ID NO1112 151-169 MMP9 ILGQEQDSFGGN SEQ ID NO1113 152-163 MMP9 ILGQEQDSFGGNFEGSQ SEQ ID NO1114 152-168 MMP9 ILGQEQDSFGGNFEGSQS SEQ ID NO1115 152-169 MMP9 LGQEQDSFGGNFEGSQ SEQ ID NO1116 153-168 MMP9 LGQEQDSFGGNFEGSQS SEQ ID NO1117 153-169 MMP9 GQEQDSFGGNFEGSQS SEQ ID NO1118 154-169 MMP9 SFGGNFEGSQS SEQ ID NO1119 159-169 MMP9 QSLVGDIGNVN SEQ ID NO1120 168-178 MMP9 INTIYLGGPFSPNV SEQ ID NO1121 189-202 MMP9 INTIYLGGPFSPNVLN SEQ ID NO1122 189-204 MMP9 IYLGGPFSPNVLN SEQ ID NO1123 192-204 MMP9 YLGGPFSPNVLN SEQ ID NO1124 193-204 MMP9 LGGPFSPN SEQ ID NO1125 194-201 MMP9 SPNVLNWRALKYEVQGEVFTKPQLWP SEQ ID NO1126 199-224 MMP9 LNWRA SEQ ID NO1127 203-207 MMP9 LNWRAL SEQ ID NO1128 203-208 MMP9 LNWRALK SEQ ID NO1129 203-209 MMP9 WRALKYE SEQ ID NO1130 205-211 MMP9 WRALKYEV SEQ ID NO1131 205-212 MMP9 WRALKYEVQGE SEQ ID NO1132 205-215 MMP9 ALKYEV SEQ ID NO1133 207-212 MMP9 LKYEVQ SEQ ID NO1134 208-213 MMP9 LKYEVQG SEQ ID NO1135 208-214 MMP9 LKYEVQGE SEQ ID NO1136 208-215 MMP9 LKYEVQGEVFTKP SEQ ID NO1137 208-220 MMP9 LKYEVQGEVFTKPQ SEQ ID NO1138 208-221 MMP9 LKYEVQGEVFTKPQLWP SEQ ID NO1139 208-224 MMP9 KYEVQGE SEQ ID NO1140 209-215 MMP9 KYEVQGEVFTKPQ SEQ ID NO1141 209-221 MMP9 KYEVQGEVFTKPQLWP SEQ ID NO1142 209-224 MMP9 YEVQGEVFTKP SEQ ID NO1143 210-220 MMP9 YEVQGEVFTKPQ SEQ ID NO1144 210-221 MMP9 YEVQGEVFTKPQLWP SEQ ID NO1145 210-224 MMP9 VQGEVFTKPQ SEQ ID NO1146 212-221 MMP9 VQGEVFTKPQLWP SEQ ID NO1147 212-224 MMP9 QGEVFTKPQ SEQ ID NO1148 213-221 MMP9 GEVFTKP SEQ ID NO1149 214-220 MMP9 GEVFTKPQ SEQ ID NO1150 214-221 MMP9 EVFTKPQ SEQ ID NO1151 215-221 MMP9 EVFTKPQLWP SEQ ID NO1152 215-224 MMP9 VFTKPQ SEQ ID NO1153 216-221 MMP9 FTKPQ SEQ ID NO1154 217-221 MMP9 FTKPQLWP SEQ ID NO1155 217-224 MMP9 TKPQLWP SEQ ID NO1156 218-224 MMP9 KPQLWP SEQ ID NO1157 219-224 MMP12 FGQTDMSRKA SEQ ID NO1036 017-026 MMP12 MSRKAFVFP SEQ ID NO1158 022-030 MMP12 MSRKAFVFPKE SEQ ID NO1159 022-032 MMP12 MSRKAFVFPKESD SEQ ID NO1160 022-034 MMP12 MSRKAFVFPKESDTS SEQ ID NO1043 022-036 MMP12 MSRKAFVFPKESDTSYVS SEQ ID NO1161 022-039 MMP12 SRKAFVFP SEQ ID NO1162 023-030 MMP12 SRKAFVFPKESD SEQ ID NO1163 023-034 MMP12 SRKAFVFPKESDTS SEQ ID NO1164 023-036 MMP12 RKAFVFP SEQ ID NO1165 024-030 MMP12 RKAFVFPKESD SEQ ID NO1166 024-034 MMP12 KAFVFP SEQ ID NO1167 025-030 MMP12 KAFVFPKE SEQ ID NO1048 025-032 MMP12 KAFVFPKESD SEQ ID NO1049 025-034 MMP12 AFVFPKE SEQ ID NO1054 026-032 MMP12 AFVFPKESDTS SEQ ID NO1168 026-036 MMP12 AFVFPKESDTSYVS SEQ ID NO1057 026-039 MMP12 FVFPKE SEQ ID NO1060 027-032 MMP12 FVFPKESD SEQ ID NO1061 027-034 MMP12 FVFPKESDTS SEQ ID NO1062 027-036 MMP12 FVFPKESDTSY SEQ ID NO1063 027-037 MMP12 FVFPKESDTSYVS SEQ ID NO1065 027-039 MMP12 VFPKESD SEQ ID NO1169 028-034 MMP12 KESDTSY SEQ ID NO1170 031-037 MMP12 KESDTSYVS SEQ ID NO1171 031-039 MMP12 VSLKAP SEQ ID NO1172 038-043 MMP12 LKAPLT SEQ ID NO1173 040-045 MMP12 LKAPLTKP SEQ ID NO1174 040-047 MMP12 YTELSSTRGYS SEQ ID NO1175 058-068 MMP12 LSSTRGYS SEQ ID NO1078 061-068 MMP12 STRGYS SEQ ID NO1080 063-068 MMP12 YATKRQDNE SEQ ID NO1176 072-080 MMP12 YATKRQDNEI SEQ ID NO1177 072-081 MMP12 YATKRQDNEIL SEQ ID NO1084 072-082 MMP12 TKRQDNEIL SEQ ID NO1178 074-082 MMP12 KRQDNEIL SEQ ID NO1179 075-082 MMP12 ILIFWSKD SEQ ID NO1180 081-088 MMP12 IFWSKD SEQ ID NO1181 083-088 MMP12 SKDIGYS SEQ ID NO1093 086-092 MMP12 WVDGKPRV SEQ ID NO1096 128-135 MMP12 WVDGKPRVR SEQ ID NO1182 128-136 MMP12 VRKSLKKGYTVGAEAS SEQ ID NO1183 135-150 MMP12 SLKKGYT SEQ ID NO1184 138-144 MMP12 SLKKGYTVG SEQ ID NO1185 138-146 MMP12 SLKKGYTVGA SEQ ID NO1186 138-147 MMP12 SLKKGYTVGAE SEQ ID NO1098 138-148 MMP12 SLKKGYTVGAEA SEQ ID NO1099 138-149 MMP12 SLKKGYTVGAEAS SEQ ID NO1100 138-150 MMP12 SLKKGYTVGAEASI SEQ ID NO1187 138-151 MMP12 LKKGYTV SEQ ID NO1101 139-145 MMP12 LKKGYTVG SEQ ID NO1102 139-146 MMP12 LKKGYTVGA SEQ ID NO1103 139-147 MMP12 LKKGYTVGAE SEQ ID NO1104 139-148 MMP12 LKKGYTVGAEA SEQ ID NO1105 139-149 MMP12 LKKGYTVGAEAS SEQ ID NO1106 139-150 MMP12 LKKGYTVGAEASI SEQ ID NO1107 139-151 MMP12 KKGYTVGAEAS SEQ ID NO1188 140-150 MMP12 KGYTVGAEAS SEQ ID NO1189 141-150 MMP12 KGYTVGAEASI SEQ ID NO1190 141-151 MMP12 SIILGQEQDSFGGN SEQ ID NO1108 150-163 MMP12 IILGQEQD SEQ ID NO1191 151-158 MMP12 IILGQEQDSFGGN SEQ ID NO1192 151-163 MMP12 IILGQEQDSFGGNFEGSQS SEQ ID NO1112 151-169 MMP12 ILGQEQDSFGGN SEQ ID NO1113 152-163 MMP12 LVGDIGNVNMWD SEQ ID NO1193 170-181 MMP12 INTIYLGGPFSPNVLN SEQ ID NO1122 189-204 MMP12 IYLGGPFSPN SEQ ID NO1194 192-201 MMP12 IYLGGPFSPNV SEQ ID NO1195 192-202 MMP12 IYLGGPFSPNVLN SEQ ID NO1123 192-204 MMP12 LGGPFSPNVLN SEQ ID NO1196 194-204 MMP12 WRALKYE SEQ ID NO1130 205-210 MMP12 YEVQGEVFTKP SEQ ID NO1143 210-220 MMP12 YEVQGEVFTKPQ SEQ ID NO1144 210-221 MMP12 YEVQGEVFTKPQLWP SEQ ID NO1145 210-224 MMP12 EVQGEVFTKP SEQ ID NO1197 211-220 MMP12 EVQGEVFTKPQLWP SEQ ID NO1198 211-224 MMP12 VQGEVFTKP SEQ ID NO1199 212-220 MMP12 VQGEVFTKPQ SEQ ID NO1146 212-221 MMP12 VQGEVFTKPQLWP SEQ ID NO1147 212-224 MMP12 GEVFTKPQLWP SEQ ID NO1200 214-224 MMP12 EVFTKP SEQ ID NO1201 215-220 MMP12 EVFTKPQLWP SEQ ID NO1152 215-224 MMP12 VFTKPQ SEQ ID NO1153 216-221 MMP12 VFTKPQL SEQ ID NO1202 216-222 MMP12 VFTKPQLWP SEQ ID NO1203 216-224 MMP12 FTKPQLWP SEQ ID NO1155 217-224 MMP12 TKPQLWP SEQ ID NO1156 218-224 MMP1 AFVFPK SEQ ID NO1033 006-031 MMP1 KAFVFPK SEQ ID NO1204 025-031 MMP1 VRKSLK SEQ ID NO1205 135-140 MMP1 YEVQGEVFTKPQLWP SEQ ID NO1145 210-224 MMP3 FGQTDMSRKA SEQ ID NO1036 017-026 MMP3 FGQTDMSRKAF SEQ ID NO1037 017-027 MMP3 MSRKAFVFPKESDTSYV SEQ ID NO1206 022-038 MMP3 MSRKAFVFPKESDTSYVS SEQ ID NO1161 022-039 MMP3 SRKAFVFPKESDTSYV SEQ ID NO1044 023-038 MMP3 SRKAFVFPKESDTSYVS SEQ ID NO1207 023-039 MMP3 RKAFVFPKESDTSYV SEQ ID NO1046 024-038 MMP3 RKAFVFPKESDTSYVS SEQ ID NO1047 024-039 MMP3 KAFVFPKE SEQ ID NO1048 025-032 MMP3 KAFVFPKESDTS SEQ ID NO1051 025-036 MMP3 KAFVFPKESDTSYVS SEQ ID NO1053 025-039 MMP3 KAFVFPKESDTSYVSL SEQ ID NO1208 025-040 MMP3 KAFVFPKESDTSYVSLK SEQ ID NO1209 025-041 MMP3 AFVFPKESDTSYVS SEQ ID NO1057 026-039 MMP3 AFVFPKESDTSYVSL SEQ ID NO1058 026-040 MMP3 AFVFPKESDTSYVSLKAP SEQ ID NO1210 026-043 MMP3 FVFPKESDTSYV SEQ ID NO1064 027-038 MMP3 FVFPKESDTSYVSLK SEQ ID NO1211 027-041 MMP3 VFPKESDTSYVSLK SEQ ID NO1212 028-041 MMP3 KESDTSYVSLKAP SEQ ID NO1213 031-043 MMP3 TKRQDNEILIFW SEQ ID NO1214 074-085 MMP3 IVEFWVDGKPRVRKS SEQ ID NO1215 124-138 MMP3 SLKKGYTVGAEA SEQ ID NO1099 138-149 MMP3 SLKKGYTVGAEAS SEQ ID NO1100 138-150 MMP3 LKKGYTVGAEA SEQ ID NO1105 139-149 MMP3 LKKGYTVGAEAS SEQ ID NO1106 139-150 MMP3 LKKGYTVGAEASI SEQ ID NO1107 139-151 MMP3 LKKGYTVGAEASII SEQ ID NO1216 139-152 MMP3 SIILGQEQDSFGGNFEGSQS SEQ ID NO1110 150-169 MMP3 IILGQEQDSFGGN SEQ ID NO1192 151-163 MMP3 IILGQEQDSFGGNFEGSQS SEQ ID NO1112 151-169 MMP3 ILGQEQDSFGGNFEGSQS SEQ ID NO1115 152-169 MMP3 LGQEQDSFGGNFEGSQS SEQ ID NO1117 153-169 MMP3 QEQDSFGGNFEGSQS SEQ ID NO1217 155-169 MMP3 SFGGNFEGSQS SEQ ID NO1119 159-169 MMP3 LVGDIGNVNMWD SEQ ID NO1193 170-181 MMP3 FVLSPDEINT SEQ ID NO1218 182-191 MMP3 YLGGPFSPNVLN SEQ ID NO1124 193-204 MMP3 LKYEVQGEVFTKPQ SEQ ID NO1138 208-221 MMP3 KYEVQGEVFTKPQ SEQ ID NO1141 209-221 MMP3 KYEVQGEVFTKPQLWP SEQ ID NO1142 209-224 MMP3 YEVQGEVFTKPQ SEQ ID NO1144 210-221 MMP3 YEVQGEVFTKPQLWP SEQ ID NO1145 210-224 MMP3 EVQGEVFTKPQLWP SEQ ID NO1198 211-224 MMP3 VQGEVFTKPQLWP SEQ ID NO1147 212-224 MMP3 GEVFTKPQLWP SEQ ID NO1200 214-224 MMP3 EVFTKPQLWP SEQ ID NO1152 215-224 MMP3 SKDIGYSFTVGGSEI SEQ ID NO1219  86-100 MMP8 FGQTDMSR SEQ ID NO1034 017-024 MMP8 FGQTDMSRK SEQ ID NO1035 017-025 MMP8 FGQTDMSRKA SEQ ID NO1036 017-026 MMP8 FGQTDMSRKAF SEQ ID NO1037 017-027 MMP8 FGQTDMSRKAFV SEQ ID NO1220 017-028 MMP8 FGQTDMSRKAFVFPKESDTSYV SEQ ID NO1040 017-038 MMP8 MSRKAFVFPKESDTSYV SEQ ID NO1206 022-038 MMP8 SRKAFVFPKESDTSYV SEQ ID NO1044 023-038 MMP8 RKAFVFPKESDTSYV SEQ ID NO1046 024-038 MMP8 KAFVFPKESDT SEQ ID NO1050 025-035 MMP8 KAFVFPKESDTS SEQ ID NO1051 025-036 MMP8 KAFVFPKESDTSYV SEQ ID NO1052 025-038 MMP8 KAFVFPKESDTSYVS SEQ ID NO1053 025-039 MMP8 AFVFPKESDTSYV SEQ ID NO1056 026-038 MMP8 FVFPKESDTSYV SEQ ID NO1064 027-038 MMP8 VFPKESDTSYV SEQ ID NO1068 028-038 MMP8 FPKESDTSYV SEQ ID NO1221 029-038 MMP8 SLKAPL SEQ ID NO1222 039-044 MMP8 SLKAPLTKP SEQ ID NO1074 039-047 MMP8 SLKAPLTKPLKA SEQ ID NO1223 039-050 MMP8 RGYSIFSYA SEQ ID NO1224 065-073 MMP8 FSYATKRQDNEILI SEQ ID NO1225 070-083 MMP8 SYATKRQDNEILI SEQ ID NO1083 071-083 MMP8 YATKRQDNEILI SEQ ID NO1085 072-083 MMP8 ATKRQDNEILI SEQ ID NO1226 073-083 MMP8 TKRQDNEILI SEQ ID NO1087 074-083 MMP8 TKRQDNEILIF SEQ ID NO1088 074-084 MMP8 FWSKDIGYS SEQ ID NO1227 084-092 MMP8 FWSKDIGYSFT SEQ ID NO1228 084-094 MMP8 FWSKDIGYSFTV SEQ ID NO1229 084-095 MMP8 WSKDIGYSFTV SEQ ID NO1230 085-095 MMP8 KSLKKGYTVGAEA SEQ ID NO1231 137-149 MMP8 SLKKGYTVGAEA SEQ ID NO1099 138-149 MMP8 LKKGYTV SEQ ID NO1101 139-145 MMP8 LKKGYTVGAEA SEQ ID NO1105 139-149 MMP8 LKKGYTVGAEAS SEQ ID NO1106 139-150 MMP8 KKGYTVGAEA SEQ ID NO1232 140-149 MMP8 GAEASIILGQE SEQ ID NO1233 146-156 MMP8 GAEASIILGQEQD SEQ ID NO1234 146-158 MMP8 SIILGQEQD SEQ ID NO1235 150-158 MMP8 SIILGQEQDSFGGNFEGSQ SEQ ID NO1109 150-168 MMP8 SIILGQEQDSFGGNFEGSQS SEQ ID NO1110 150-169 MMP8 IILGQEQDSFGGN SEQ ID NO1192 151-163 MMP8 IILGQEQDSFGGNFEGSQ SEQ ID NO1236 151-168 MMP8 IILGQEQDSFGGNFEGSQS SEQ ID NO1112 151-169 MMP8 ILGQEQDSFGGN SEQ ID NO1113 152-163 MMP8 ILGQEQDSFGGNFEGS SEQ ID NO1237 152-167 MMP8 ILGQEQDSFGGNFEGSQ SEQ ID NO1114 152-168 MMP8 ILGQEQDSFGGNFEGSQS SEQ ID NO1115 152-169 MMP8 LGQEQDSFGGN SEQ ID NO1238 153-163 MMP8 LGQEQDSFGGNFEGS SEQ ID NO1239 153-167 MMP8 LGQEQDSFGGNFEGSQ SEQ ID NO1116 153-168 MMP8 LGQEQDSFGGNFEGSQS SEQ ID NO1117 153-169 MMP8 LGQEQDSFGGNFEGSQSL SEQ ID NO1240 153-170 MMP8 LGQEQDSFGGNFEGSQSLV SEQ ID NO1241 153-171 MMP8 QDSFGGNFEGSQS SEQ ID NO1242 157-169 MMP8 SFGGNFEGSQ SEQ ID NO1243 159-168 MMP8 SFGGNFEGSQS SEQ ID NO1119 159-169 MMP8 SFGGNFEGSQSLV SEQ ID NO1244 159-171 MMP8 LVGDIGNVNMW SEQ ID NO1245 170-180 MMP8 INTIYLGGPFSPN SEQ ID NO1246 189-201 MMP8 TIYLGGPFSPN SEQ ID NO1247 191-201 MMP8 IYLGGPFSPN SEQ ID NO1194 192-201 MMP8 YLGGPFSPNV SEQ ID NO1248 193-202 MMP8 YLGGPFSPNVLN SEQ ID NO1124 193-204 MMP8 LGGPFSPNVLN SEQ ID NO1196 194-204 MMP8 VLNWRA SEQ ID NO1249 202-207 MMP8 VLNWRAL SEQ ID NO1250 202-208 MMP8 VLNWRALK SEQ ID NO1251 202-209 MMP8 LNWRAL SEQ ID NO1128 203-208 MMP8 LNWRALK SEQ ID NO1129 203-209 MMP8 LNWRALKYEV SEQ ID NO1252 203-212 MMP8 NWRAL SEQ ID NO1253 204-208 MMP8 NWRALKY SEQ ID NO1254 204-210 MMP8 NWRALKYEV SEQ ID NO1255 204-212 MMP8 NWRALKYEVQ SEQ ID NO1256 204-213 MMP8 WRALKYE SEQ ID NO1130 205-211 MMP8 WRALKYEVQ SEQ ID NO1257 205-213 MMP8 WRALKYEVQGE SEQ ID NO1132 205-215 MMP8 RALKYEV SEQ ID NO1258 206-212 MMP8 RALKYEVQ SEQ ID NO1259 206-213 MMP8 RALKYEVQGE SEQ ID NO1260 206-215 MMP8 ALKYEV SEQ ID NO1133 207-212 MMP8 ALKYEVQGEVFTKPQ SEQ ID NO1261 207-221 MMP8 LKYEVQGE SEQ ID NO1136 208-215 MMP8 LKYEVQGEVFTKPQ SEQ ID NO1138 208-221 MMP8 KYEVQGEVFTKPQ SEQ ID NO1141 209-221 MMP8 KYEVQGEVFTKPQLWP SEQ ID NO1142 209-224 MMP8 YEVQGEVFTKPQ SEQ ID NO1144 210-221 MMP8 YEVQGEVFTKPQLWP SEQ ID NO1145 210-224 MMP8 EVQGEVFTKPQ SEQ ID NO1262 211-221 MMP8 EVQGEVFTKPQLWP SEQ ID NO1198 211-224 MMP8 VQGEVFTKPQ SEQ ID NO1146 212-221 MMP8 VQGEVFTKPQLWP SEQ ID NO1147 212-224 MMP8 QGEVFTKPQ SEQ ID NO1148 213-221 MMP8 QGEVFTKPQL SEQ ID NO1263 213-222 MMP8 QGEVFTKPQLWP SEQ ID NO1264 213-224 MMP8 GEVFTKPQ SEQ ID NO1150 214-221 MMP8 GEVFTKPQLWP SEQ ID NO1200 214-224 MMP8 VFTKPQ SEQ ID NO1153 216-221 MMP8 VFTKPQLWP SEQ ID NO1203 216-224 MMP8 FTKPQLWP SEQ ID NO1155 217-224 MMP8 TKPQLWP SEQ ID NO1156 218-224 ADAMTS-1 ESDTSYVSLK SEQ ID NO1265 032-041 ADAMTS-1 QEQDSFGGNFEGSQ SEQ ID NO1266 155-168 ADAMTS-1 QEQDSFGGNFEGSQSLVG SEQ ID NO1267 155-172 ADAMTS-1 GNFEGSQSLVG SEQ ID NO1268 162-172 ADAMTS-1 YEVQGEVFT SEQ ID NO1269 210-218 ADAMTS-1 YEVQGEVFTKPQ SEQ ID NO1144 210-221 ADAMTS-1 GEVFTKPQ SEQ ID NO1150 214-221 ADAMTS-8 VFPKESDTSYVS SEQ ID NO1069 028-039 ADAMTS-8 QEQDSFGGNFEGSQSLVG SEQ ID NO1267 155-172 ADAMTS-8 EINTIYL SEQ ID NO1270 188-194 ADAMTS-8 KYEVQ SEQ ID NO1271 209-213 ADAMTS-8 KYEVQGE SEQ ID NO1140 209-215 Cat K FGQTDMSR SEQ ID NO1034 017-024 Cat K AFVFPK SEQ ID NO1033 026-031 Cat K FVFPK SEQ ID NO1059 027-031 Cat K ESDTSYVSLK SEQ ID NO1265 032-041 Cat K ESDTSYVSLKAPLT SEQ ID NO1272 032-045 Cat K SDTSYVSLK SEQ ID NO1273 033-041 Cat K DTSYVSLK SEQ ID NO1274 034-041 Cat K STRGYS SEQ ID NO1080 063-068 Cat K IFWSKDIG SEQ ID NO1275 083-090 Cat K KGYTVGAE SEQ ID NO1276 141-148 Cat K AEASIILGQEQDSFG SEQ ID NO1277 147-161 Cat K LGQEQDSFG SEQ ID NO1278 153-161 Cat K LGQEQDSFGGNFE SEQ ID NO1279 153-165 Cat K GQEQDSFG SEQ ID NO1280 154-161 Cat K GQEQDSFGGNFE SEQ ID NO1281 154-165 Cat K GQEQDSFGGNFEGSQ SEQ ID NO1282 154-168 Cat K GQEQDSFGGNFEGSQS SEQ ID NO1118 154-169 Cat K QEQDSFGGN SEQ ID NO1283 155-163 Cat K QEQDSFGGNFE SEQ ID NO1284 155-165 Cat K QEQDSFGGNFEG SEQ ID NO1285 155-166 Cat K QEQDSFGGNFEGS SEQ ID NO1286 155-167 Cat K QEQDSFGGNFEGSQ SEQ ID NO1266 155-168 Cat K QEQDSFGGNFEGSQS SEQ ID NO1217 155-169 Cat K GNFEGSQSLV SEQ ID NO1287 162-171 Cat K GNFEGSQSLVG SEQ ID NO1268 162-172 Cat K GNFEGSQSLVGDIG SEQ ID NO1288 162-175 Cat K GSQSLVGDIG SEQ ID NO1289 166-175 Cat K GSQSLVGDIGNVN SEQ ID NO1290 166-178 Cat K DFVLSPDEIN SEQ ID NO1291 181-190 Cat K FVLSPDEINT SEQ ID NO1218 182-191 Cat K VLSPDEINT SEQ ID NO1291 183-191 Cat K GPFSPNVLN SEQ ID NO1292 196-204 Cat K SPNVLNWR SEQ ID NO1293 199-206 Cat K KYEVQG SEQ ID NO1294 209-214 Cat K YEVQGEVFT SEQ ID NO1269 210-218 Cat K YEVQGEVFTKPQ SEQ ID NO1144 210-221 Cat K VQGEVFTKPQ SEQ ID NO1146 212-221 Cat K GEVFTKPQ SEQ ID NO1150 214-221 Cat K EVFTKPQ SEQ ID NO1151 215-221 Cat S FGQTDMSR SEQ ID NO1034 017-024 Cat S AFVFPKESDTSYVS SEQ ID NO1057 026-039 Cat S FVFPKESDTSYVS SEQ ID NO1065 027-039 Cat S VFPKESDTSYVS SEQ ID NO1069 028-039 Cat S FPKESDTSYVS SEQ ID NO1071 029-039 Cat S ESDTSYVSLK SEQ ID NO1265 032-041 Cat S TSWESASGIVE SEQ ID NO1295 116-126 Cat S KGYTVG SEQ ID NO1296 141-146 Cat S QEQDSFGGNFE SEQ ID NO1284 155-165 Cat S QEQDSFGGNFEG SEQ ID NO1285 155-166 Cat S QEQDSFGGNFEGSQ SEQ ID NO1266 155-168 Cat S QEQDSFGGNFEGSQS SEQ ID NO1217 155-169 Cat S QEQDSFGGNFEGSQSLV SEQ ID NO1297 155-171 Cat S QEQDSFGGNFEGSQSLVG SEQ ID NO1267 155-172 Cat S SFGGNFEGSQSLVG SEQ ID NO1298 159-172 Cat S GNFEGSQSLVG SEQ ID NO1268 162-172 Cat S GNFEGSQSLVGDIG SEQ ID NO1288 162-175 Cat S SPDEINTIYL SEQ ID NO1299 185-194 Cat S SPDEINTIYLG SEQ ID NO1300 185-195 Cat S LGGPFSPNVLN SEQ ID NO1196 194-204 Cat S GGPFSPNVLN SEQ ID NO1301 195-204 Cat S GPFSPNVLN SEQ ID NO1292 196-204 Cat S ALKYE SEQ ID NO1302 207-211 Cat S ALKYEVQ SEQ ID NO1303 207-213 Cat S YEVQGEVF SEQ ID NO1304 210-217 Cat S YEVQGEVFT SEQ ID NO1269 210-218 Cat S YEVQGEVFTKPQ SEQ ID NO1144 210-221 Cat S YEVQGEVFTKPQLWP SEQ ID NO1145 210-224 Cat S VQGEVFTKPQLWP SEQ ID NO1147 212-224 Cat S GEVFTKPQ SEQ ID NO1150 214-221 Cat S GEVFTKPQLWP SEQ ID NO1200 214-224 Cat S EVFTKPQLWP SEQ ID NO1152 215-224 Cat S TKPQLWP SEQ ID NO1156 218-224 Cat S KPQLWP SEQ ID NO1157 219-224 *numbers in the sequence of CRP

Accordingly, in a method of the invention, said peptide fragments preferably comprise a neo-epitope formed by cleavage of CRP by a protease at a site marked by the sign * in any one of the above partial sequences of CRP in Table 20 or at either end of any partial sequence of CRP in Table 21.

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of CRP.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 22 N-terminal sequences of protease generated peptide fragments of CRP. CRP AFVFPK SEQ ID NO1033 VSLKAP SEQ ID NO1172 TDMSRK SEQ ID NO1307 VFPKES SEQ ID NO1310 LSSTRG SEQ ID NO1313 KRQDNE SEQ ID NO1315 SIILGQ SEQ ID NO1318 IYLGGP SEQ ID NO1321 ALKYEV SEQ ID NO1133 KPQLWP SEQ ID NO1157 LVGDIG SEQ ID NO1327 GAEASI SEQ ID NO1330 EINTIY SEQ ID NO1333 TSWESA SEQ ID NO1336 YEVQGE SEQ ID NO1339 RKAFVF SEQ ID NO1342 TKPQLW SEQ ID NO1345 SDTSYV SEQ ID NO1347 IFSYAT SEQ ID NO1348 EFWVDG SEQ ID NO1351 LGQEQD SEQ ID NO1354 SPNVLN SEQ ID NO1357 QGEVFT SEQ ID NO1358 IFWSKD SEQ ID NO1181 FSYATK SEQ ID NO1361 VLNWRA SEQ ID NO1249 GSQSLV SEQ ID NO1365 LKYEVQ SEQ ID NO1134 GNFEGS SEQ ID NO1369 FVFPKE SEQ ID NO1060 FYTELS SEQ ID NO1374 SFGGNF SEQ ID NO1305 KAFVFP SEQ ID NO1167 MSRKAF SEQ ID NO1308 FPKESD SEQ ID NO1311 SSTRGY SEQ ID NO1314 WSKDIG SEQ ID NO1316 IILGQE SEQ ID NO1319 YLGGPF SEQ ID NO1322 KYEVQG SEQ ID NO1294 YTELSS SEQ ID NO1325 QEQDSF SEQ ID NO1328 QDSFGG SEQ ID NO1331 DTSYVS SEQ ID NO1334 SPDEIN SEQ ID NO1337 FVLSPD SEQ ID NO1340 IVEFWV SEQ ID NO1343 EVQGEV SEQ ID NO1346 SLKAPL SEQ ID NO1222 SYATKR SEQ ID NO1349 WVDGKP SEQ ID NO1352 GQEQDS SEQ ID NO1355 LNWRA SEQ ID NO1127 GEVFTK SEQ ID NO1359 VRKSLK SEQ ID NO1205 ATKRQD SEQ ID NO1362 NWRAL SEQ ID NO1253 DFVLSP SEQ ID NO1366 TKRQDN SEQ ID NO1368 SLKKGY SEQ ID NO1370 INTIYL SEQ ID NO1372 WRALKY SEQ ID NO1375 FGQTDM SEQ ID NO1306 EVFTKP SEQ ID NO1201 SRKAFV SEQ ID NO1309 KESDTS SEQ ID NO1312 STRGYS SEQ ID NO1080 SKDIGY SEQ ID NO1317 ILGQEQ SEQ ID NO1320 LGGPFS SEQ ID NO1323 VQGEVF SEQ ID NO1324 ILIFWS SEQ ID NO1326 RGYSIF SEQ ID NO1329 TIYLGG SEQ ID NO1332 AEASII SEQ ID NO1335 GGPFSP SEQ ID NO1338 LKKGYT SEQ ID NO1341 ESDTSY SEQ ID NO1344 FVFPK SEQ ID NO1059 LKAPLT SEQ ID NO1173 YATKRQ SEQ ID NO1350 VDGKPR SEQ ID NO1353 QSLVGD SEQ ID NO1356 LNWRAL SEQ ID NO1128 VFTKPQ SEQ ID NO1153 KKGYTV SEQ ID NO1360 FWSKDI SEQ ID NO1363 NWRALK SEQ ID NO1364 VLSPDE SEQ ID NO1367 KGYTVG SEQ ID NO1296 KSLKKG SEQ ID NO1371 RALKYE SEQ ID NO1373 GPFSPN SEQ ID NO1376 or with any of the following sequences at the C-terminal of a peptide:

TABLE 23 C-terminal sequences of protease generated peptide fragments of CRP. CRP AFVFPK SEQ ID NO1033 SPDEIN SEQ ID NO1337 KESDTS SEQ ID NO1312 LTKPLK SEQ ID NO1379 KDIGYS SEQ ID NO1380 GAEASI SEQ ID NO1330 SPNVLN SEQ ID NO1357 ALKYEV SEQ ID NO1133 KAFVFP SEQ ID NO1167 LKKGYT SEQ ID NO1341 PRVRKS SEQ ID NO1384 GYSFTV SEQ ID NO1386 INTIYL SEQ ID NO1372 NVLNWR SEQ ID NO1389 VGAEAS SEQ ID NO1392 QTDMSR SEQ ID NO1394 PKESDT SEQ ID NO1395 DNEILI SEQ ID NO1397 YTVGAE SEQ ID NO1400 FEGSQS SEQ ID NO1403 LNWRA SEQ ID NO1127 KYEVQG SEQ ID NO1294 KRQDNE SEQ ID NO1315 FTKPQL SEQ ID NO1406 SLKAPL SEQ ID NO1222 GSQSLV SEQ ID NO1365 FGGNFE SEQ ID NO1412 VQGEVF SEQ ID NO1324 DMSRKA SEQ ID NO1415 FPKESD SEQ ID NO1311 KPQLWP SEQ ID NO1157 DSFGGN SEQ ID NO1378 SDTSYV SEQ ID NO1347 STRGYS SEQ ID NO1080 DGKPRV SEQ ID NO1381 QGEVFT SEQ ID NO1358 GPFSPN SEQ ID NO1376 YEVQGE SEQ ID NO1339 VSLKAP SEQ ID NO1172 LGQEQD SEQ ID NO1354 TVGSEI SEQ ID NO1385 IILGQE SEQ ID NO1319 WSKDIG SEQ ID NO1316 ASGIVE SEQ ID NO1390 APLTKP SEQ ID NO1393 TDMSRK SEQ ID NO1307 TSYVSL SEQ ID NO1396 QDNEIL SEQ ID NO1398 TVGAEA SEQ ID NO1401 GNFEGS SEQ ID NO1369 LNWRAL SEQ ID NO1128 EVFTKP SEQ ID NO1201 RQDNEI SEQ ID NO1405 VRKSLK SEQ ID NO1205 TKPLKA SEQ ID NO1408 GNVNMW SEQ ID NO1410 GGNFEG SEQ ID NO1413 GKPRVR SEQ ID NO1414 EILIEW SEQ ID NO1416 IGYSFT SEQ ID NO1417 PDEINT SEQ ID NO1377 VFTKPQ SEQ ID NO1153 DTSYVS SEQ ID NO1334 YATKRQ SEQ ID NO1350 VDGKPR SEQ ID NO1353 NFEGSQ SEQ ID NO1382 RALKYE SEQ ID NO1373 LKYEVQ SEQ ID NO1134 LKAPLT SEQ ID NO1173 NVNMWD SEQ ID NO1383 SRKAFV SEQ ID NO1309 EGSQSL SEQ ID NO1387 EQDSFG SEQ ID NO1388 NTIYLG SEQ ID NO1391 FVFPKE SEQ ID NO1060 MSRKAF SEQ ID NO1308 ESDTSY SEQ ID NO1344 NEILIF SEQ ID NO1399 PFSPNV SEQ ID NO1402 DIGNVN SEQ ID NO1404 NWRALK SEQ ID NO1364 VFTKPQ SEQ ID NO1153 IFWSKD SEQ ID NO1181 SYVSLK SEQ ID NO1407 SIFSYA SEQ ID NO1409 SQSLVG SEQ ID NO1411 LVGDIG SEQ ID NO1327 FWSKDI 1363 SEQ ID NO KKGYTV SEQ ID NO1360

Elastin

Several candidate proteases may be responsible for the digestion of elastin in fibrotic tissue We have through a range of in vitro cleavages of pure native proteins determined that the enzymes listed in the following table cleaved elastin at least at the cleavage sites at each end of the following sequences or at the cleavage sites marked ‘.’ or where no ‘.’ is shown, at the ends of the sequences:

TABLE 24 Elastin fragments generated by specific proteases. Protease Sequence between cleavage sites Nos* MMP9 + 12 GVPGAIPGGVPG SEQ ID NO1418 028-039 MMP9 + 12 AIPGGVPGGVFYPGAGLG SEQ ID NO1419 032-049 MMP9 + 12 AIPGGVPGGVFYPGAGLGA SEQ ID NO1420 032-050 MMP9 + 12 GVPGGVFYPGAGLGA SEQ ID NO1421 036-050 MMP9 + 12 GVPGGVFYPGAGLGALG SEQ ID NO1422 036-052 MMP9 + 12 VPGGVFYPGAGLGALGG SEQ ID NO1423 037-053 MMP9 + 12 GVFYPGAGLGALGGGALGPGG SEQ ID NO1424 040-060 MMP9 + 12 VFYPGAGLG SEQ ID NO1425 041-049 MMP9 + 12 VFYPGAGLGA SEQ ID NO1426 041-050 MMP9 + 12 VFYPGAGLGAL SEQ ID NO1427 041-051 MMP9 + 12 VFYPGAGLGALG SEQ ID NO1428 041-052 MMP9 + 12 VFYPGAGLGALGG SEQ ID NO1429 041-053 MMP9 + 12 VFYPGAGLGALGGG SEQ ID NO1430 041-054 MMP9 + 12 VFYPGAGLGALGGGAL SEQ ID NO1431 041-056 MMP9 + 12 VFYPGAGLGALGGGALG SEQ ID NO1432 041-057 MMP9 + 12 VFYPGAGLGALGGGALGPG SEQ ID NO1433 041-059 MMP9 + 12 VFYPGAGLGALGGGALGPGG SEQ ID NO1434 041-060 MMP9 + 12 VFYPGAGLGALGGGALGPGGKPLKPVPGG SEQ ID NO1435 041-069 MMP9 + 12 LGALGGGALGPGGKPLKPVPGG SEQ ID NO1436 048-069 MMP9 + 12 ALGGGALGPGGKPLKPVPGG SEQ ID NO1437 050-069 MMP9 + 12 LGGGALGPGGKPLKPVPG SEQ ID NO1438 051-068 MMP9 + 12 LGGGALGPGGKPLKPVPGG SEQ ID NO1439 051-069 MMP9 + 12 GGALGPGGKPLKPVPGG SEQ ID NO1440 053-069 MMP9 + 12 LGPGGKPLKPVPGG SEQ ID NO1441 056-069 MMP9 + 12 GPGGKPLKPVPGG SEQ ID NO1442 057-069 MMP9 + 12 PGGKPLKPVPGG SEQ ID NO1443 058-069 MMP9 + 12 GKPLKPVPGG SEQ ID NO1444 060-069 MMP9 + 12 PLKPVPGG SEQ ID NO1445 062-069 MMP9 + 12 LKPVPGG SEQ ID NO 063-069 SEQ ID NO1446 MMP9 + 12 GLAGAGLGAGLGAFP SEQ ID NO1447 069-083 MMP9 + 12 GLAGAGLGAGLGAFPA SEQ ID NO1448 069-084 MMP9 + 12 LAGAGLGAGLG SEQ ID NO1449 070-080 MMP9 + 12 LAGAGLGAGLGAFP SEQ ID NO1450 070-083 MMP9 + 12 LAGAGLGAGLGAFPA SEQ ID NO1451 070-084 MMP9 + 12 LAGAGLGAGLGAFPAVT SEQ ID NO1452 070-086 MMP9 + 12 LAGAGLGAGLGAFPAVTFPG SEQ ID NO1453 070-089 MMP9 + 12 LAGAGLGAGLGAFPAVTFPGA SEQ ID NO1454 070-090 MMP9 + 12 LAGAGLGAGLGAFPAVTFPGALVPGG SEQ ID NO1455 070-095 MMP9 + 12 LAGAGLGAGLGAFPAVTFPGALVPGGVA SEQ ID NO1456 070-097 MMP9 + 12 LAGAGLGAGLGAFPAVTFPGALVPGGVADAAAA SEQ ID NO1457 070-102 MMP9 + 12 AGAGLGAGLGAFPAVTFPGALVPGG SEQ ID NO1458 071-095 MMP9 + 12 GAGLGAGLGAFPA SEQ ID NO1459 072-084 MMP9 + 12 GAGLGAGLGAFPAVTFPGA SEQ ID NO1460 072-090 MMP9 + 12 AGLGAGLGAFPA SEQ ID NO1461 073-084 MMP9 + 12 GLGAGLGAFPA SEQ ID NO1462 074-084 MMP9 + 12 LGAGLGAFPA SEQ ID NO1463 075-084 MMP9 + 12 LGAGLGAFPAVTFPGA SEQ ID NO1464 075-090 MMP9 + 12 LGAGLGAFPAVTFPGALVPGG SEQ ID NO1465 075-095 MMP9 + 12 LGAGLGAFPAVTFPGALVPGGVADAAAA SEQ ID NO1466 075-102 MMP9 + 12 AGLGAFPAVTFPG SEQ ID NO1467 077-089 MMP9 + 12 LGAFPAVTFPGA SEQ ID NO1468 079-090 MMP9 + 12 LGAFPAVTFPGALVPGGVA SEQ ID NO1469 079-097 MMP9 + 12 LGAFPAVTFPGALVPGGVADAAAA SEQ ID NO1470 079-102 MMP9 + 12 AFPAVTFPGALVPGG SEQ ID NO1471 081-095 MMP9 + 12 AVTFPGALVPGG SEQ ID NO1472 084-095 MMP9 + 12 AVTFPGALVPGGVADAAAA SEQ ID NO1473 084-102 MMP9 + 12 VTFPGALVPGG SEQ ID NO1474 085-095 MMP9 + 12 VTFPGALVPGGVADAAAA SEQ ID NO1475 085-102 MMP9 + 12 LVPGGVADAAAA SEQ ID NO1476 091-102 MMP9 + 12 LVPGGVADAAAAYK SEQ ID NO1477 091-104 MMP9 + 12 VADAAAAYK SEQ ID NO1478 096-104 MMP9 + 12 KAAKAGA SEQ ID NO1479 104-110 MMP9 + 12 LGVSAGAVVPQPGA SEQ ID NO1480 121-134 MMP9 + 12 VPGVGLPGVYPGGVLPGAR SEQ ID NO1481 141-159 MMP9 + 12 PGVGLPGVYPGGVLPGAR SEQ ID NO1482 142-159 MMP9 + 12 GLPGVYPGGVLPGAR SEQ ID NO1483 145-159 MMP9 + 12 PGVYPGGVLPGAR SEQ ID NO1484 147-159 MMP9 + 12 ARFPGVG SEQ ID NO1485 158-164 MMP9 + 12 ARFPGVGVLPG SEQ ID NO1486 158-168 MMP9 + 12 RFPGVGVLPGVPTGAG SEQ ID NO1487 159-174 MMP9 + 12 FPGVGVLPGVPTG SEQ ID NO1488 160-172 MMP9 + 12 FPGVGVLPGVPTGA SEQ ID NO1489 160-173 MMP9 + 12 FPGVGVLPGVPTGAGV SEQ ID NO1490 160-175 MMP9 + 12 FPGVGVLPGVPTGAGVKPK SEQ ID NO1491 160-178 MMP9 + 12 KPKAPGV SEQ ID NO1492 176-182 MMP9 + 12 PKAPGV SEQ ID NO1493 177-182 MMP9 + 12 GAFAGIPGVGPFG SEQ ID NO1494 184-196 MMP9 + 12 VGPFGGPQPGVPLGYP SEQ ID NO1495 192-207 MMP9 + 12 GPQPGVPLGYP SEQ ID NO1496 197-207 MMP9 + 12 PQPGVPLGYP SEQ ID NO1497 198-207 MMP9 + 12 PGVPLGYP SEQ ID NO1498 200-207 MMP9 + 12 GYPIKAPK SEQ ID NO1499 205-212 MMP9 + 12 PKLPGGY SEQ ID NO1500 211-217 MMP9 + 12 YTTGKLPYGYGPG SEQ ID NO1501 221-233 MMP9 + 12 YTTGKLPYGYGPGGVAGAAGK SEQ ID NO1502 221-241 MMP9 + 12 TTGKLPYGYG SEQ ID NO1503 222-231 MMP9 + 12 TTGKLPYGYGPGGVAGAAGK SEQ ID NO1504 222-241 MMP9 + 12 LPYGYGPGGVAGAAGK SEQ ID NO1505 226-241 MMP9 + 12 GYGPGGVAGAAGK SEQ ID NO1506 229-241 MMP9 + 12 YGPGGVAGAAGK SEQ ID NO1507 230-241 MMP9 + 12 AGYPTGTGVGPQAAAAAAAK SEQ ID NO1508 242-261 MMP9 + 12 TGVGPQAAAAAAAK SEQ ID NO1509 248-261 MMP9 + 12 PQAAAAAAAK SEQ ID NO1510 252-261 MMP9 + 12 FGAGAAGVLPGVGGAGVPGVPGAIPGIGG SEQ ID NO1511 266-294 MMP9 + 12 FGAGAAGVLPGVGGAGVPGVPGAIPGIGGIAGVGTPAA SEQ ID NO1512 266-303 MMP9 + 12 GVLPGVGGAGVPGVPG SEQ ID NO1513 272-287 MMP9 + 12 VLPGVGGAGVPGVPGAIPGIGG SEQ ID NO1514 273-294 MMP9 + 12 VLPGVGGAGVPGVPGAIPGIGGIAGVGTPA SEQ ID NO1515 273-302 MMP9 + 12 VLPGVGGAGVPGVPGAIPGIGGIAGVGTPAA SEQ ID NO1516 273-303 MMP9 + 12 GAG VPG VPGAIPG SEQ ID NO1517 279-291 MMP9 + 12 GAGVPGVPGAIPGIGGIAGVG SEQ ID NO1518 279-299 MMP9 + 12 AGVPGVPGAIPGIG SEQ ID NO1519 280-293 MMP9 + 12 AGVPGVPGAIPGIGG SEQ ID NO1520 280-294 MMP9 + 12 AGVPGVPGAIPGIGGIAG SEQ ID NO1521 280-297 MMP9 + 12 AGVPGVPGAIPGIGGIAGVGTPA SEQ ID NO1522 280-302 MMP9 + 12 GVPGVPGAIPGIGG SEQ ID NO1523 281-294 MMP9 + 12 GVPGVPGAIPGIGGIA SEQ ID NO1524 281-296 MMP9 + 12 GVPGVPGAIPGIGGIAGVG SEQ ID NO1525 281-299 MMP9 + 12 VPGVPGAIPGIGG SEQ ID NO1526 282-294 MMP9 + 12 GVPGAIPGIGGIAGVGTPA SEQ ID NO1527 284-302 MMP9 + 12 VPGAIPGIGGIAGVG SEQ ID NO1528 285-299 MMP9 + 12 VPGAIPGIGGIAGVGTPA SEQ ID NO1529 285-302 MMP9 + 12 VPGAIPGIGGIAGVGTPAAA SEQ ID NO1530 285-304 MMP9 + 12 VPGAIPGIGGIAGVGTPAAAAAAAAAAK SEQ ID NO1531 285-312 MMP9 + 12 AIPGIGGIAGVG SEQ ID NO1532 288-299 MMP9 + 12 AIPGIGGIAGVGTPA SEQ ID NO1533 288-302 MMP9 + 12 AIPGIGGIAGVGTPAA SEQ ID NO1534 288-303 MMP9 + 12 AIPGIGGIAGVGTPAAA SEQ ID NO1535 288-304 MMP9 + 12 AIPGIGGIAGVGTPAAAAAA SEQ ID NO1536 288-307 MMP9 + 12 AIPGIGGIAGVGTPAAAAAAAAAAK SEQ ID NO1537 288-312 MMP9 + 12 IPGIGGIAGVGTPAAA SEQ ID NO1538 289-304 MMP9 + 12 IGGIAGVGTPAAAA SEQ ID NO1539 292-305 MMP9 + 12 GIAGVGTPAAAA SEQ ID NO154O 294-305 MMP9 + 12 GIAGVGTPAAAAAAAA SEQ ID NO1541 294-309 MMP9 + 12 GIAGVGTPAAAAAAAAAAK SEQ ID NO1542 294-312 MMP9 + 12 IAGVGTPAAAAAAAA SEQ ID NO1543 295-309 MMP9 + 12 IAGVGTPAAAAAAAAA SEQ ID NO1544 295-310 MMP9 + 12 IAGVGTPAAAAAAAAAAK SEQ ID NO1545 295-312 MMP9 + 12 TPAAAAAAAAAAK SEQ ID NO1546 300-312 MMP9 + 12 PAAAAAAAAAAK SEQ ID NO1547 301-312 MMP9 + 12 AAAAAAAAAAK SEQ ID NO1548 302-312 MMP9 + 12 AAAAAAAAAK SEQ ID NO1549 303-312 MMP9 + 12 AAAAAAAAK SEQ ID NO1550 304-312 MMP9 + 12 AAAAAAAAKA SEQ ID NO1551 304-313 MMP9 + 12 AAAAAAAK SEQ ID NO1552 305-312 MMP9 + 12 LVPGGPGFGPGVVGVPGA SEQ ID NO1553 322-339 MMP9 + 12 GPGFGPGVVGVPG SEQ ID NO1554 326-338 MMP9 + 12 GPGFGPGVVGVPGAGVPGVG SEQ ID NO1555 326-345 MMP9 + 12 GPGFGPGVVGVPGAGVPGVGVPGAGIPVVPG SEQ ID NO1556 326-356 MMP9 + 12 PGFGPGVVGVPG SEQ ID NO1557 327-338 MMP9 + 12 PGFGPGVVGVPGA SEQ ID NO1558 327-339 MMP9 + 12 PGFGPGVVGVPGAG SEQ ID NO1559 327-340 MMP9 + 12 PGVVGVPGAGVPG SEQ ID NO1560 331-343 MMP9 + 12 PGVVGVPGAGVPGVGVPG SEQ ID NO1561 331-348 MMP9 + 12 PGVVGVPGAGVPGVGVPGAGIPVVPGA SEQ ID NO1562 331-357 MMP9 + 12 VVGVPGAGVPGVGVPGA SEQ ID NO1563 333-349 MMP9 + 12 VGVPGAGVPGVGVPGAGIPVVPGAGIPGAAVPGVVSPEA SEQ ID NO1564 334-372 MMP9 + 12 AGVPGVGVPGAGIPVVPG SEQ ID NO1565 339-356 MMP9 + 12 GVPGVGVPGAGIPVVPG SEQ ID NO1566 340-356 MMP9 + 12 GVPGVGVPGAGIPVVPGA SEQ ID NO1567 340-357 MMP9 + 12 VPGVGVPGAGIPVVPG SEQ ID NO1568 341-356 MMP9 + 12 VGVPGAGIPVVPG SEQ ID NO1569 344-356 MMP9 + 12 VGVPGAGIPVVPGAGIPG SEQ ID NO1570 344-361 MMP9 + 12 VPGAGIPVVPG SEQ ID NO1571 346-356 MMP9 + 12 AGIPVVPGAGIPG SEQ ID NO1572 349-361 MMP9 + 12 AGIPVVPGAGIPGAAVPGVVSPEAAAK SEQ ID NO1573 349-375 MMP9 + 12 GIPVVPGAGIPG SEQ ID NO1574 350-361 MMP9 + 12 IPGAAVPGVVSPEAAAK SEQ ID NO1575 359-375 MMP9 + 12 GAAVPGVVSPEAAAK SEQ ID NO1576 361-375 MMP9 + 12 AVPGVVSPEAAAK SEQ ID NO1577 363-375 MMP9 + 12 VPGVVSPEAAAK SEQ ID NO1578 364-375 MMP9 + 12 YGARPGVG SEQ ID NO1579 383-390 MMP9 + 12 YGARPGVGVG SEQ ID NO1580 383-392 MMP9 + 12 YGARPGVGVGGIPT SEQ ID NO1581 383-396 MMP9 + 12 YGARPGVGVGGIPTY SEQ ID NO1582 383-397 MMP9 + 12 YGARPGVGVGGIPTYG SEQ ID NO1583 383-398 MMP9 + 12 YGARPGVGVGGIPTYGVG SEQ ID NO1584 383-400 MMP9 + 12 YGARPGVGVGGIPTYGVGA SEQ ID NO1585 383-401 MMP9 + 12 YGARPGVGVGGIPTYGVGAG SEQ ID NO1586 383-402 MMP9 + 12 GARPGVGV SEQ ID NO1587 384-391 MMP9 + 12 GARPGVGVGG SEQ ID NO1588 384-393 MMP9 + 12 GARPGVGVGGIP SEQ ID NO1589 384-395 MMP9 + 12 GARPGVGVGGIPTY SEQ ID NO1590 384-397 MMP9 + 12 GARPGVGVGGIPTYGV SEQ ID NO1591 384-399 MMP9 + 12 GARPGVGVGGIPTYGVG SEQ ID NO1592 384-400 MMP9 + 12 GARPGVGVGGIPTYGVGAGGF SEQ ID NO1593 384-404 MMP9 + 12 GARPGVGVGGIPTYGVGAGGFPGF SEQ ID NO1594 384-407 MMP9 + 12 GARPGVGVGGIPTYGVGAGGFPGFG SEQ ID NO1595 384-408 MMP9 + 12 GARPGVGVGGIPTYGVGAGGFPGFGVGVG SEQ ID NO1596 384-412 MMP9 + 12 ARPGVGVGG SEQ ID NO1597 385-393 MMP9 + 12 ARPGVGVGGIP SEQ ID NO1598 385-395 MMP9 + 12 ARPGVGVGGIPTY SEQ ID NO1599 385-397 MMP9 + 12 ARPGVGVGGIPTYGVGA SEQ ID NO1600 385-401 MMP9 + 12 ARPGVGVGGIPTYGVGAGG SEQ ID NO1601 385-403 MMP9 + 12 ARPGVGVGGIPTYGVGAGGFPG SEQ ID NO1602 385-406 MMP9 + 12 ARPGVGVGGIPTYGVGAGGFPGF SEQ ID NO1603 385-407 MMP9 + 12 RPGVGVG SEQ ID NO1604 386-392 MMP9 + 12 RPGVGVGG SEQ ID NO1605 386-393 MMP9 + 12 PGVGVGGIPTY SEQ ID NO1606 387-397 MMP9 + 12 PGVGVGGIPTYG SEQ ID NO1607 387-398 MMP9 + 12 PGVGVGGIPTYGVGAG SEQ ID NO1608 387-412 MMP9 + 12 VGGIPTYGVGAG SEQ ID NO1609 391-402 MMP9 + 12 GVGAGGFPGFGVGVGGIPGVA SEQ ID NO1610 398-418 MMP9 + 12 VGAGGFPGFGVGVG SEQ ID NO1611 399-412 MMP9 + 12 VGVGGIPGVAGVPSVGGVPGVGGVPGVGISPEA SEQ ID NO1612 409-441 MMP9 + 12 VAGVPSVGGVPGVGGVPG SEQ ID NO1613 417-434 MMP9 + 12 VAGVPSVGGVPGVGGVPGVGISPEA SEQ ID NO1614 417-441 MMP9 + 12 SVGGVPGVGGVPGVGISPEA SEQ ID NO1615 422-441 MMP9 + 12 VGGVPGVGGVPGVGISPEA SEQ ID NO1616 423-441 MMP9 + 12 GVPGVGGVPGVGIS SEQ ID NO1617 425-438 MMP9 + 12 GVPGVGGVPGVGIS PEA SEQ ID NO1618 425-441 MMP9 + 12 GVPGVGGVPGVGISPEAQA SEQ ID NO1619 425-443 MMP9 + 12 GVPGVGISPEAQAAAAAK SEQ ID NO1620 431-448 MMP9 + 12 GVGTPAAAAAK SEQ ID NO1621 482-492 MMP9 + 12 TPAAAAAK SEQ ID NO1622 485-492 MMP9 + 12 FGLVPGVGVAPGVG SEQ ID NO1623 500-513 MMP9 + 12 FGLVPGVGVAPGVGVAPG SEQ ID NO1624 500-517 MMP9 + 12 FGLVPGVGVAPGVGVAPGVGVAPG SEQ ID NO1625 500-523 MMP9 + 12 FGLVPGVGVAPGVGVAPGVGVAPGVG SEQ ID NO1626 500-525 MMP9 + 12 FGLVPGVGVAPGVGVAPGVGVAPGVGLAPG SEQ ID NO1627 500-529 MMP9 + 12 FGLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO1628 500-535 MMP9 + 12 FGLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPGV SEQ ID NO1629 500-536 MMP9 + 12 FGLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPGVGVAPG SEQ ID NO1630 500-541 MMP9 + 12 GLVPGVGVAPG SEQ ID NO1631 501-511 MMP9 + 12 GLVPGVGVAPGV SEQ ID NO1632 501-512 MMP9 + 12 GLVPGVGVAPGVGVA SEQ ID NO1633 501-515 MMP9 + 12 GLVPGVGVAPGVGVAP SEQ ID NO1634 501-516 MMP9 + 12 GLVPGVGVAPGVGVAPG SEQ ID NO1635 501-517 MMP9 + 12 GLVPGVGVAPGVGVAPGVG SEQ ID NO1636 501-519 MMP9 + 12 GLVPGVGVAPGVGVAPGVGVAPG SEQ ID NO1637 501-523 MMP9 + 12 GLVPGVGVAPGVGVAPGVGVAPGVGL SEQ ID NO1638 501-524 MMP9 + 12 GLVPGVGVAPGVGVAPGVGVAPGVGLA SEQ ID NO1639 501-525 MMP9 + 12 GLVPGVGVAPGVGVAPGVGVAPGVGLAPG SEQ ID NO1640 501-527 MMP9 + 12 GLVPGVGVAPGVGVAPGVGVAPGVGLAPGVG SEQ ID NO1641 501-529 MMP9 + 12 GLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVA SEQ ID NO1642 501-531 MMP9 + 12 GLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO1643 501-533 MMP9 + 12 LVPGVGVAPGVG SEQ ID NO1644 502-513 MMP9 + 12 LVPGVGVAPGVGVAPG SEQ ID NO1645 502-517 MMP9 + 12 LVPGVGVAPGVGVAPGVG SEQ ID NO1646 502-519 MMP9 + 12 LVPGVGVAPGVGVAPGVGVAPGVG SEQ ID NO1647 502-525 MMP9 + 12 LVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPGVG SEQ ID NO1648 502-537 MMP9 + 12 PGVGVAPGVGVAPG SEQ ID NO1649 504-517 MMP9 + 12 VGVAPGVGVAPGVGV SEQ ID NO1650 506-520 MMP9 + 12 VGVAPGVGVAPGVGVAPGVG SEQ ID NO1651 506-525 MMP9 + 12 VGVAPGVGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO1652 506-535 MMP9 + 12 VAPGVGVAPGVGVAPG SEQ ID NO1653 508-523 MMP9 + 12 VAPGVGVAPGVGVAPGVG SEQ ID NO1654 508-525 MMP9 + 12 VAPGVGVAPGVGVAPGVGLAPGVG SEQ ID NO1655 508-531 MMP9 + 12 VAPGVGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO1656 508-535 MMP9 + 12 VGVAPGVGVAPGVGLA SEQ ID NO1657 512-527 MMP9 + 12 VGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO1658 512-535 MMP9 + 12 VGVAPGVGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1659 512-552 MMP9 + 12 VAPGVGVAPGVGLAPGVGVAPGVG SEQ ID NO1660 514-537 MMP9 + 12 VAPGVGVAPGVGLAPGVGVAPGVGVA SEQ ID NO1661 514-539 MMP9 + 12 VAPGVGVAPGVGLAPGVGVAPGVGVAPG SEQ ID NO1662 514-541 MMP9 + 12 VAPGVGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGP SEQ ID NO1663 514-550 MMP9 + 12 VAPGVGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1664 514-552 MMP9 + 12 PGVGVAPGVGLAPG SEQ ID NO1665 516-529 MMP9 + 12 PGVGVAPGVGLAPGVGVAP SEQ ID NO1666 516-534 MMP9 + 12 PGVGVAPGVGLAPGVGVAPGVG SEQ ID NO1667 516-537 MMP9 + 12 VGVAPGVGLAPGVGVA SEQ ID NO1668 518-533 MMP9 + 12 VGVAPGVGLAPGVGVAP SEQ ID NO1669 518-534 MMP9 + 12 VGVAPGVGLAPGVGVAPGVGVAPG SEQ ID NO1670 518-541 MMP9 + 12 VGVAPGVGLAPGVGVAPGVGVAPGVG SEQ ID NO1671 518-543 MMP9 + 12 VGVAPGVGLAPGVGVAPGVGVAPGVGVAPG SEQ ID NO1672 518-547 MMP9 + 12 VGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1673 518-552 MMP9 + 12 GVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1674 519-552 MMP9 + 12 VAPGVGLAPGVGVA SEQ ID NO1675 520-533 MMP9 + 12 VAPGVGLAPGVGVAPG SEQ ID NO1676 520-535 MMP9 + 12 VAPGVGLAPGVGVAPGVG SEQ ID NO1677 520-537 MMP9 + 12 VAPGVGLAPGVGVAPGVGVA SEQ ID NO1678 520-539 MMP9 + 12 VAPGVGLAPGVGVAPGVGVAPG SEQ ID NO1679 520-541 MMP9 + 12 VAPGVGLAPGVGVAPGVGVAPGVGVA SEQ ID NO1680 520-545 MMP9 + 12 VAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1681 520-552 MMP9 + 12 PGVGLAPGVGVAPG SEQ ID NO1682 522-535 MMP9 + 12 GVGLAPGVGVAPGVGVAPG SEQ ID NO1683 523-541 MMP9 + 12 VGLAPGVGVAPGVG SEQ ID NO1684 524-537 MMP9 + 12 VGLAPGVGVAPGVGVAPG SEQ ID NO1685 524-541 MMP9 + 12 VGLAPGVGVAPGVGVAPGVGVAPGIG SEQ ID NO1686 524-549 MMP9 + 12 VGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1687 524-552 MMP9 + 12 VGLAPGVGVAPGVGVAPGVGVAPGIGPG SEQ ID NO1688 524-553 MMP9 + 12 VGLAPGVGVAPGVGVAPGVGVAPGIGPGGVAAA SEQ ID NO1689 524-556 MMP9 + 12 LAPGVGVAPGVGVAPGVG SEQ ID NO1690 526-543 MMP9 + 12 LAPGVGVAPGVGVAPGVGVA SEQ ID NO1691 526-545 MMP9 + 12 LAPGVGVAPGVGVAPGVGVAPGIGP SEQ ID NO1692 526-550 MMP9 + 12 LAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1693 526-552 MMP9 + 12 GVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1694 529-552 MMP9 + 12 VGVAPGVGVAPGVGVA SEQ ID NO1695 530-545 MMP9 + 12 VGVAPGVGVAPGVGVAPG SEQ ID NO1696 530-547 MMP9 + 12 VGVAPGVGVAPGVGVAPGIGPG SEQ ID NO1697 530-551 MMP9 + 12 VGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO1698 530-552 MMP9 + 12 VGVAPGVGVAPGVGVAPGIGPGGVAAA SEQ ID NO1699 530-556 MMP9 + 12 VAPGVGVAPGVGVAP SEQ ID NO1700 532-546 MMP9 + 12 VAPGVGVAPGVGVAPGIG SEQ ID NO1701 532-549 MMP9 + 12 VAPGVGVAPGVGVAPGIGPGG SEQ ID NO1702 532-552 MMP9 + 12 PGVGVAPGVGVAPGIGPG SEQ ID NO1703 534-551 MMP9 + 12 PGVGVAPGVGVAPGIGPGG SEQ ID NO1704 534-552 MMP9 + 12 VGVAPGVGVAPGIGPGG SEQ ID NO1705 536-552 MMP9 + 12 VGVAPGVGVAPGIGPGGVAA SEQ ID NO1706 536-555 MMP9 + 12 VAPGVGVAPGIGPG SEQ ID NO1707 538-551 MMP9 + 12 PGVGVAPGIGPG SEQ ID NO1708 540-551 MMP9 + 12 VGVAPGIGPGGVAA SEQ ID NO1709 542-555 MMP9 + 12 PGGVAAAAK SEQ ID NO1710 550-558 MMP9 + 12 LRAAAGL SEQ ID NO1711 569-575 MMP9 + 12 LRAAAGLG SEQ ID NO1712 569-576 MMP9 + 12 LRAAAGLGA SEQ ID NO1713 569-577 MMP9 + 12 AAAGLGAGIPGLGVG SEQ ID NO1714 571-585 MMP9 + 12 AAAGLGAGIPGLGVGVG SEQ ID NO1715 571-587 MMP9 + 12 LGAGIPGLGVG SEQ ID NO1716 575-585 MMP9 + 12 LGAGIPGLGVGVG SEQ ID NO1717 575-587 MMP9 + 12 LGAGIPGLGVGVGVPGLGVG SEQ ID NO1718 575-594 MMP9 + 12 LGAGIPGLGVGVGVPG SEQ ID NO1719 575-590 MMP9 + 12 LGAGIPGLGVGVGVPGLGVGA SEQ ID NO1720 575-595 MMP9 + 12 LGAGIPGLGVGVGVPGL SEQ ID NO1721 575-591 MMP9 + 12 LGAGIPGLGVGVGVPGLG SEQ ID NO1722 575-592 MMP9 + 12 LGAGIPGLGVGVGVPGLGVGAGVPG SEQ ID NO1723 575-599 MMP9 + 12 LGAGIPGLGVGVGVPGLGVGAGVPGLG SEQ ID NO1724 575-601 MMP9 + 12 LGAGIPGLGVGVGVPGLGVGAGVPGLGVG SEQ ID NO1725 575-603 MMP9 + 12 LGAGIPGLGVGVGVPGLGVGAGVPGLGVGAGVPGFG SEQ ID NO1726 575-610 MMP9 + 12 GAGIPGLGVGVGVPGLG SEQ ID NO1727 576-592 MMP9 + 12 AGIPGLGVGVGVPG SEQ ID NO1728 577-590 MMP9 + 12 GIPGLGVGVGVPGLGVGA SEQ ID NO1729 578-595 MMP9 + 12 LGVGVGVPGLGVGA SEQ ID NO1730 582-595 MMP9 + 12 VGVPGLGVGAGVPG SEQ ID NO1731 586-599 MMP9 + 12 VGVPGLGVGAGVPGL SEQ ID NO1732 586-600 MMP9 + 12 VGVPGLGVGAGVPGLG SEQ ID NO1733 586-601 MMP9 + 12 VGVPGLGVGAGVPGLGVG SEQ ID NO1734 586-603 MMP9 + 12 VGVPGLGVGAGVPGLGVGA SEQ ID NO1735 586-604 MMP9 + 12 VGAGVPGLGVGAGVPGFG SEQ ID NO1736 593-610 MMP9 + 12 PGALAAAK SEQ ID NO1737 646-653 MMP9 + 12 AKYGAAVPGVLGGLGA SEQ ID NO1738 655-670 MMP9 + 12 YGAAVPGVLGG SEQ ID NO1739 657-667 MMP9 + 12 YGAAVPGVLGGLG SEQ ID NO1740 657-669 MMP9 + 12 YGAAVPGVLGGLGA SEQ ID NO1741 657-670 MMP9 + 12 YGAAVPGVLGGLGALG SEQ ID NO1742 657-672 MMP9 + 12 YGAAVPGVLGGLGALGGVGIPGG SEQ ID NO1743 657-679 MMP9 + 12 YGAAVPGVLGGLGALGGVGIPGGVVGAGPAA SEQ ID NO1744 657-687 MMP9 + 12 GAAVPGVLGGLG SEQ ID NO1745 658-669 MMP9 + 12 GAAVPGVLGGLGALGGVGIPGG SEQ ID NO1746 658-679 MMP9 + 12 AVPGVLGGLGA SEQ ID NO1747 660-670 MMP9 + 12 AVPGVLGGLGALGGVGIPGG SEQ ID NO1748 660-679 MMP9 + 12 VLGGLGALGGVGIPGG SEQ ID NO1749 664-679 MMP9 + 12 GGLGALGGVGIPGGVVGAGPA SEQ ID NO1750 666-686 MMP9 + 12 GGLGALGGVGIPGGVVGAGPAAA SEQ ID NO1751 666-688 MMP9 + 12 LGALGGVGIPGG SEQ ID NO1752 668-379 MMP9 + 12 LGALGGVGIPGGVVGAGPA SEQ ID NO1753 668-686 MMP9 + 12 LGALGGVGIPGGVVGAGPAA SEQ ID NO1754 668-687 MMP9 + 12 LGALGGVGIPGGVVGAGPAAA SEQ ID NO1755 668-688 MMP9 + 12 LGALGGVGIPGGVVGAGPAAAA SEQ ID NO1756 668-689 MMP9 + 12 ALGGVGIPGGVVGAGPAA SEQ ID NO1757 670-687 MMP9 + 12 ALGGVGIPGGVVGAGPAAA SEQ ID NO1758 670-688 MMP9 + 12 LGGVGIPGGV SEQ ID NO1759 671-680 MMP9 + 12 LGGVGIPGGVVGAGPA SEQ ID NO1760 671-686 MMP9 + 12 LGGVGIPGGVVGAGPAAA SEQ ID NO1761 671-688 MMP9 + 12 LGGVGIPGGVVGAGPAAAAA SEQ ID NO1762 671-690 MMP9 + 12 LGGVGIPGGVVGAGPAAAAAAAK SEQ ID NO1763 671-693 MMP9 + 12 GVGIPGGVVGAGPAAAA SEQ ID NO1764 673-689 MMP9 + 12 GVGIPGGVVGAGPAAAAAAAK SEQ ID NO1765 673-693 MMP9 + 12 VGIPGGVVGAGPAAA SEQ ID NO1766 674-688 MMP9 + 12 VGIPGGVVGAGPAAAAAAAK SEQ ID NO1767 674-693 MMP9 + 12 IPGGVVGAGPAAAA SEQ ID NO1768 676-689 MMP9 + 12 VVGAGPAAAAAAAK SEQ ID NO1769 680-693 MMP9 + 12 VGAGPAAAAAAAK SEQ ID NO1770 681-693 MMP9 + 12 AGPAAAAAAAK SEQ ID NO1771 683-693 MMP9 + 12 GPAAAAAAAK SEQ ID NO1772 684-693 MMP9 + 12 PAAAAAAAK SEQ ID NO1773 685-693 MMP9 + 12 FGLVGAAGLGGLGVGGLGVPGVGG SEQ ID NO1774 701-724 MMP9 + 12 GLVGAAGLGGLG SEQ ID NO1775 702-713 MMP9 + 12 GLVGAAGLGGLGVGG SEQ ID NO1776 702-716 MMP9 + 12 GLVGAAGLGGLGVGGLGVPGVG SEQ ID NO1777 702-723 MMP9 + 12 GLVGAAGLGGLGVGGLGVPGVGG SEQ ID NO1778 702-724 MMP9 + 12 LVGAAGLGGLGVG SEQ ID NO1779 703-715 MMP9 + 12 LVGAAGLGGLGVGG SEQ ID NO1780 703-716 MMP9 + 12 LVGAAGLGGLGVGGL SEQ ID NO1781 703-717 MMP9 + 12 LVGAAGLGGLGVGGLGVPGVGGLG SEQ ID NO1782 703-726 MMP9 + 12 LVGAAGLGGLGVGGLGVPGVGGLGGIPPAAA SEQ ID NO1783 703-733 MMP9 + 12 VGAAGLGGLGVGG SEQ ID NO1784 704-716 MMP9 + 12 LGGLGVGGLGVPG SEQ ID NO1785 709-721 MMP9 + 12 LGGLGVGGLGVPGVG SEQ ID NO1786 709-723 MMP9 + 12 LGGLGVGGLGVPGVGGL SEQ ID NO1787 709-725 MMP9 + 12 LGGLGVGGLGVPGVGGLG SEQ ID NO1788 709-726 MMP9 + 12 LGVGGLGVPGVGGLG SEQ ID NO1789 712-726 MMP9 + 12 GLGVPGVGGLGGIPPAAAAK SEQ ID NO1790 716-735 MMP9 + 12 LGGIPPAAAAK SEQ ID NO1791 725-735 MMP9 + 12 LGGVLGGAGQFPL SEQ ID NO1792 744-756 MMP9 + 12 LGGVLGGAGQFPLGGVAAR SEQ ID NO1793 744-762 MMP9 + 12 LGGVLGGAGQFPLGGVAARPG SEQ ID NO1794 744-764 MMP9 + 12 LGGVLGGAGQFPLGGVAARPGFG SEQ ID NO1795 744-766 MMP9 + 12 GGVLGGAGQFPLGGVAARPG SEQ ID NO1796 745-764 MMP9 + 12 GAGQFPLGGVAAR SEQ ID NO1797 750-762 MMP9 + 12 GAGQFPLGGVAARPGFG SEQ ID NO1798 750-766 MMP9 + 12 AGQFPLGGVAARPGFG SEQ ID NO1799 751-766 MMP9 + 12 FPLGGVAARPG SEQ ID NO1800 754-764 MMP9 + 12 PLGGVAAR SEQ ID NO1801 755-762 MMP9 + 12 PLGGVAARPG SEQ ID NO1802 755-764 MMP9 + 12 PLGGVAARPGFG SEQ ID NO1803 755-766 MMP9 + 12 PLGGVAARPGFGL SEQ ID NO1804 755-767 MMP9 + 12 PLGGVAARPGFGLSPIFPG SEQ ID NO1805 755-773 MMP9 + 12 LGGVAAR SEQ ID NO1806 756-762 MMP9 + 12 LGGVAARP SEQ ID NO1807 756-763 MMP9 + 12 LGGVAARPG SEQ ID NO1808 756-764 MMP9 + 12 LGGVAARPGF SEQ ID NO1809 756-765 MMP9 + 12 LGGVAARPGFG SEQ ID NO1810 756-766 MMP9 + 12 LGGVAARPGFGL SEQ ID NO1811 756-767 MMP9 + 12 LGGVAARPGFGLSP SEQ ID NO1812 756-769 MMP9 + 12 LGGVAARPGFGLSPIFPG SEQ ID NO1813 756-773 MMP9 + 12 LGGVAARPGFGLSPIFPGG SEQ ID NO1814 756-774 MMP9 + 12 LGGVAARPGFGLSPIFPGGA SEQ ID NO1815 756-775 MMP9 + 12 GGVAARPGFG SEQ ID NO1816 757-766 MMP9 + 12 GGVAARPGFGL SEQ ID NO1817 757-767 MMP9 + 12 GGVAARPGFGLSPIFPGGA SEQ ID NO1818 757-775 MMP9 + 12 GVAARPGFGLSPIF SEQ ID NO1819 758-771 MMP9 + 12 GVAARPGFGLSPIFP SEQ ID NO1820 758-772 MMP9 + 12 VAARPGFG SEQ ID NO1821 759-766 MMP9 + 12 VAARPGFGLSPIFP SEQ ID NO1822 759-772 MMP9 + 12 VAARPGFGLSPIFPG SEQ ID NO1823 759-773 MMP9 + 12 RPGFGLSPIFPG SEQ ID NO1824 762-773 MMP9 + 12 PGFGLSPIFPGG SEQ ID NO1825 763-774 MMP9 + 12 PGFGLSPIFPGGA SEQ ID NO1826 763-775 ADAMTS-1 P.GVGLPGVYPGGVLPGAR.F SEQ ID NO1827 143-159 ADAMTS-1 G.VGLPGVYPGGVLPGAR.F SEQ ID NO1828 144-159 ADAMTS-1 G.LPGVYPGGVLPGAR.F SEQ ID NO1829 146-159 ADAMTS-1 P.GVYPGGVLPGAR.F SEQ ID NO1830 148-159 ADAMTS-1 K.AGYPTGTGVGPQAAAAAAAK.A SEQ ID NO1831 242-261 ADAMTS-1 G.GPGFGPGVVGVPGAGVPGVGVPGA.G SEQ ID NO1832 326-349 ADAMTS-1 G.FGPGVVGVPGAGVPGVGVPG.A SEQ ID NO1833 329-348 ADAMTS-1 F.GPGVVGVPGAGVPGVGVPG.A SEQ ID NO1834 330-348 ADAMTS-1 G.VPGVGVPGAGIPVVPG.A SEQ ID NO1835 341-356 ADAMTS-1 G.ARPGVGVGGIPTYGVG.A SEQ ID NO1836 385-400 ADAMTS-1 G.ARPGVGVGGIPTYGVGAGG.F SEQ ID NO1837 385-403 ADAMTS-1 A.RPGVGVGGIPTYGVGAG.G SEQ ID NO1838 386-402 ADAMTS-1 G.GVPGVGGVPGVGISPEAQAAAA.A SEQ ID NO1839 425-446 ADAMTS-1 G.VPGVGISPEAQAAAAAK.A SEQ ID NO1840 432-448 ADAMTS-1 G.VGISPEAQAAAAAK.A SEQ ID NO1841 435-448 ADAMTS-1 V.PGVGVAPGVGVAPGVGVAPGVGL.A SEQ ID NO1842 504-526 ADAMTS-1 G.VAPGVGVAPGVGVAPGVGLAPGVGVAPG.V SEQ ID NO1843 508-535 ADAMTS-1 G.VGVAPGVGVAPGVGLAPGVG.V SEQ ID NO1844 512-531 ADAMTS-1 G.VGVAPGVGVAPGVGLAPGVGVAPGVG.V SEQ ID NO1845 512-537 ADAMTS-1 A.PGVGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1846 528-551 ADAMTS-1 G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1847 532-551 ADAMTS-1 G.AAVPGVLGGLGALGGVGIPG.G SEQ ID NO1848 659-678 ADAMTS-1 G.AAGLGGLGVGGLGVPGVGGLG.G SEQ ID NO1849 706-726 ADAMTS-4 P.GVGLPGVYPGGVLPGAR.F SEQ ID NO1827 143-159 ADAMTS-4 G.LPGVYPGGVLPGAR.F SEQ ID NO1829 146-159 ADAMTS-4 K.AGYPTGTGVGPQAAAAAAAK.A SEQ ID NO1831 242-261 ADAMTS-4 G.GAGVPGVPGAIPGIGGIAGVG.T SEQ ID NO1850 279-299 ADAMTS-4 G.AGVPGVPGAIPGIGGIAGVG.T SEQ ID NO1851 280-299 ADAMTS-4 A.GVGTPAAAAAAAAAAK.A SEQ ID NO1852 297-312 ADAMTS-4 G.VGTPAAAAAAAAAAK.A SEQ ID NO1853 298-312 ADAMTS-4 G.GPGFGPGVVGVPGAGVPGVGVPG.A SEQ ID NO1854 326-348 ADAMTS-4 G.ARPGVGVGGIPTYGVGA.G SEQ ID NO1855 385-401 ADAMTS-4 A.RPGVGVGGIPTYGVGAG.G SEQ ID NO1838 386-402 ADAMTS-4 A.RPGVGVGGIPTYGVGAGG.F SEQ ID NO1856 386-403 ADAMTS-4 G.VGISPEAQAAAAAK.A SEQ ID NO1841 435-448 ADAMTS-4 G.VGVAPGVGVAPGVGVAPGVGLAPGVG.V SEQ ID NO1857 506-531 ADAMTS-4 A.PGVGVAPGVGLAPGVGVAPGVGVA.P SEQ ID NO1858 516-539 ADAMTS-4 G.VGVAPGVGLAPGVGVAPGVG.V SEQ ID NO1859 518-537 ADAMTS-4 L.APGVGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1860 527-551 ADAMTS-4 Y.GAAVPGVLGGLGALGGVGIPG.G SEQ ID NO1861 658-678 ADAMTS-4 G.AAVPGVLGGLGALGGVGIPG.G SEQ ID NO1848 659-678 ADAMTS-4 G.GAGQFPLGGVAARPGFGL.S SEQ ID NO1862 750-767 ADAMTS-8 L.VPGGVADAAAAYK.A SEQ ID NO1863 092-104 ADAMTS-8 G.VGLPGVYPGGVLPGAR.F SEQ ID NO1828 144-159 ADAMTS-8 G.LPGVYPGGVLPGAR.F SEQ ID NO1829 146-159 ADAMTS-8 P.GVYPGGVLPGAR.F SEQ ID NO1830 148-159 ADAMTS-8 V.YPGGVLPGAR.F SEQ ID NO1864 150-159 ADAMTS-8 F.GPGVVGVPGAGVPGVGVPG.A SEQ ID NO1834 330-348 ADAMTS-8 G.ARPGVGVGGIPTYGVGA.G SEQ ID NO1855 385-401 ADAMTS-8 V.APGVGVAPGVGVAPGVGLAPGVGV.A SEQ ID NO1865 509-532 ADAMTS-8 L.APGVGVAPGVGVAPGVGV.A SEQ ID NO1866 527-544 ADAMTS-8 L.APGVGVAPGVGVAPGVGVAPG.I SEQ ID NO1867 527-547 ADAMTS-8 L.APGVGVAPGVGVAPGVGVAPGIG.P SEQ ID NO1868 527-549 ADAMTS-8 L.APGVGVAPGVGVAPGVGVAPGIGP.G SEQ ID NO1869 527-550 ADAMTS-8 L.APGVGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1860 527-551 ADAMTS-8 L.APGVGVAPGVGVAPGVGVAPGIGPGGVAA.A SEQ ID NO1870 527-555 ADAMTS-8 G.VGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1871 530-551 ADAMTS-8 G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1847 532-551 ADAMTS-8 G.AAVPGVLGGLGALGGVGIPG.G SEQ ID NO1848 659-678 ADAMTS-8 G.AAVPGVLGGLGALGGVGIPGG.V SEQ ID NO1872 659-679 ADAMTS-8 A.AVPGVLGGLGALGGVGIPG.G SEQ ID NO1873 660-678 ADAMTS-8 A.VPGVLGGLGALGGVGIPGG.V SEQ ID NO1874 661-679 ADAMTS-8 A.GQFPLGGVAARPGFGL.S SEQ ID NO1875 752-767 Cat K G.ALVPGGVADAAAAYK.A SEQ ID NO1876 090-104 Cat K G.LPYTTGKLPYGYGPG.G SEQ ID NO1877 219-233 Cat K A.AAAAAAKAAAKFGA.G SEQ ID NO1878 255-268 Cat K A.GVGTPAAAAAAAAAAK.A SEQ ID NO1852 297-312 Cat K A.AAAAAAAAAKAAKYGA.A SEQ ID NO1879 303-318 Cat K G.FGPGVVGVPGAGVPGVGVPG.A SEQ ID NO1833 329-348 Cat K G.VGISPEAQAAAAAK.A SEQ ID NO1841 435-448 Cat K G.VAPGVGVAPGVGVAPGVGLAPGVG.V SEQ ID NO1880 508-531 Cat K G.VGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1871 530-551 Cat K G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1847 532-551 Cat S T.FPGALVPGGVADAAAAYK.A SEQ ID NO1881 087-104 Cat S G.VGLPGVYPGGVLPGAR.F SEQ ID NO1828 144-159 Cat S G.LPGVYPGGVLPGARFPGVG.V SEQ ID NO1882 146-164 Cat S G.YPTGTGVGPQAAAAAAAK.A SEQ ID NO1883 244-261 Cat S G.GAGVPGVPGAIPGIGGIAGVG.T SEQ ID NO1850 279-299 Cat S G.TPAAAAAAAAAAKAAK.Y SEQ ID NO1884 300-315 Cat S G.VPGAGVPGVGVPGAGIPVVP.G SEQ ID NO1885 336-355 Cat S G.VPGAGVPGVGVPGAGIPVVPGAGIPG.A SEQ ID NO1886 336-361 Cat S G.ISPEAQAAAAAKAAK.Y SEQ ID NO1887 437-451 Cat S V.PGVGVAPGVGVAPGVGVA.P SEQ ID NO1888 504-521 Cat S G.VAPGVGVAPGVGVAPGIGPGGVA.A SEQ ID NO1889 532-554 Cat S G.IPGGVVGAGPAAAAAAAK.A SEQ ID NO1890 676-693 MMP1 G.GVLPGARFPGVGVLPGVPTGA.G SEQ ID NO1891 153-173 MMP1 G.GVPGVGGVPGVGISPEA.Q SEQ ID NO1892 425-441 MMP1 V.PGVGVAPGVGVAPGVGVA.P SEQ ID NO1888 504-521 MMP1 G.VGVAPGVGVAPGVGVAPGVG.L SEQ ID NO1893 506-525 MMP1 G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1847 532-551 MMP1 A.AVPGVLGGLGALGGVGIPG.G SEQ ID NO1873 660-678 MMP1 MMP3 G.ALVPGGVADAAAAYK.A SEQ ID NO1876 090-104 MMP3 G.YPTGTGVGPQAAAAAAAK.A SEQ ID NO1883 244-261 MMP3 G.VPGVPGAIPGIGGIAGVG.T SEQ ID NO1894 282-299 MMP3 F.GPGVVGVPGAGVPGVGVPGA.G SEQ ID NO1895 330-349 MMP3 G.VGISPEAQAAAAAK.A SEQ ID NO1841 435-448 MMP3 G.VGVAPGVGVAPGVGLAPGVG.V SEQ ID NO1844 512-531 MMP3 G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO1847 532-551 MMP8 P.GVYPGGVLPGAR.F SEQ ID NO1830 148-159 MMP8 K.AGYPTGTGVGPQAAAAAAAK.A SEQ ID NO1831 242-261 MMP8 G.VPGVPGAIPGIGGIAGVG.T SEQ ID NO1894 282-299 MMP8 F.GPGVVGVPGAGVPGVGVPG.A SEQ ID NO1834 330-348 MMP8 G.VPGVGVPGAGIPVVPGA.G SEQ ID NO1896 341-357 MMP8 G.ARPGVGVGGIPTYGVG.A SEQ ID NO1836 385-400 MMP8 A.RPGVGVGGIPTYGVGAG.G SEQ ID NO1838 386-402 MMP8 G.VGVAPGVGVAPGVGVAP.G SEQ ID NO1897 506-522 MMP8 G.VGVAPGVGVAPGVGLAPGVG.V SEQ ID NO1844 512-531 MMP8 G.VGVAPGVGVAPGVGVAP.G SEQ ID NO1897 530-546 MMP8 G.IPGGVVGAGPAAAAAAAK.A SEQ ID NO1890 676-693 *Aminoacid residue numbers in the human elastin sequence

Accordingly, in a method of the invention, said peptide fragments preferably comprise a neo-epitope formed by cleavage of elastin by a protease at an N- or C-terminal site, or where indicated a site marked by the sign in any one of the partial sequences of elastin in Table 24.

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of elastin.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 25 N-terminal sequences of protease generated peptide fragments of elastin. Elastin GVPGAI SEQ ID NO1898 ALGGGA SEQ ID NO1901 PLKPVP SEQ ID NO1904 GLGAGL SEQ ID NO1907 LVPGGV SEQ ID NO1910 PVGYPG SEQ ID NO1913 VGPFGG SEQ ID NO1916 TTGKLP SEQ ID NO1919 FGAGAA SEQ ID NO1922 AGIPGL SEQ ID NO1925 TPAAAA SEQ ID NO1928 VGVPGA SEQ ID NO1931 GIPVVP SEQ ID NO1934 ARPGVG SEQ ID NO1937 SVGGVP SEQ ID NO1940 VGVAPG SEQ ID NO1943 GVPVAP SEQ ID NO1946 LGAGIP SEQ ID NO1949 PGALAA SEQ ID NO1952 ALGGVG SEQ ID NO1955 AGPAAA SEQ ID NO1958 GLGVPG SEQ ID NO1961 PLGGVA SEQ ID NO1964 GVGLPG SEQ ID NO1967 VPGVPV SEQ ID NO1970 APGVGV SEQ ID NO1973 GPGFGP SEQ ID NO1976 VPGGVF SEQ ID NO1979 LGPGGK SEQ ID NO1982 LAGAGL SEQ ID NO1985 LGAFPA SEQ ID NO1988 LGVSAG SEQ ID NO1991 FPGVGV SEQ ID NO1994 PGVPLG SEQ ID NO1996 YGPGGV SEQ ID NO1999 GAGVPG SEQ ID NO2002 IPGIGG SEQ ID NO2005 VPGVGV SEQ ID NO2008 AVPGVV SEQ ID NO2011 GVGAGG SEQ ID NO2014 FGLVPG SEQ ID NO2017 PGVGLA SEQ ID NO2020 LRAAAG SEQ ID NO2023 GIPGLG SEQ ID NO2026 AAAGLG SEQ ID NO2029 VGIPGG SEQ ID NO2032 GLVGAA SEQ ID NO2035 GGVLGG SEQ ID NO2038 GVAARP SEQ ID NO2041 GVYPGG SEQ ID NO2044 AAVPGV SEQ ID NO2047 VPGVLG SEQ ID NO2050 ISPEAQ SEQ ID NO2053 LGALGG SEQ ID NO2056 GKPLKP SEQ ID NO2059 AGLGAG SEQ ID NO2062 VTFPGA SEQ ID NO2065 GLPGVY SEQ ID NO2068 GAFAGI SEQ ID NO2071 YTTGKL SEQ ID NO2074 PQAAAA SEQ ID NO2077 AIPGGV SEQ ID NO1899 LGGGAL SEQ ID NO1902 LKPVPG SEQ ID NO1905 LGAGLG SEQ ID NO1908 VADAAA SEQ ID NO1911 ARFPGV SEQ ID NO1914 GPQPGV SEQ ID NO1917 LPYGYG SEQ ID NO1920 GVLPGV SEQ ID NO1923 VPGAIP SEQ ID NO1926 PAAAAA SEQ ID NO1929 AVGPGV SEQ ID NO1932 IPGAAV SEQ ID NO1935 RPGVGV SEQ ID NO1938 VGGVPG SEQ ID NO1941 VAPGVG SEQ ID NO1944 GAGIPG SEQ ID NO1947 PGFGPG SEQ ID NO1950 AKYGAA SEQ ID NO1953 LGGVGI SEQ ID NO1956 GPAAAA SEQ ID NO1959 LGGIPP SEQ ID NO1962 LGGVAA SEQ ID NO1965 VGLPGV SEQ ID NO1968 VPGVGI SEQ ID NO1971 VPGGVA SEQ ID NO1974 YPTGTG SEQ ID NO1977 GVFYPG SEQ ID NO1980 GPGGKP SEQ ID NO1983 AGAGLG SEQ ID NO1986 AFPAVT SEQ ID NO1989 VPGVGL SEQ ID NO1992 KPGAPG SEQ ID NO1995 GYPIKA SEQ ID NO1997 AGYPTG SEQ ID NO2000 AGVPGV SEQ ID NO2003 IGGIAG SEQ ID NO2006 VPVGVA SEQ ID NO2009 VPGVVS SEQ ID NO2012 VGAGGF SEQ ID NO2015 GLVPGV SEQ ID NO2018 GVGLAP SEQ ID NO2021 LVGAAG SEQ ID NO2024 LGVGVG SEQ ID NO2027 AVPGVL SEQ ID NO2030 IPGGVV SEQ ID NO2033 VGAAGL SEQ ID NO2036 GAGQFP SEQ ID NO2039 VAARPG SEQ ID NO2042 LPYTTG SEQ ID NO2045 AAGLGG SEQ ID NO2048 GQFPLG SEQ ID NO2051 GVLPGA SEQ ID NO2054 VPGVPG SEQ ID NO2057 IAGVGT SEQ ID NO2060 VVGVPG SEQ ID NO2063 AGIPVV SEQ ID NO2066 GARPGV SEQ ID NO2069 VAGVPS SEQ ID NO2072 PGVGVA SEQ ID NO2075 LAPGVG SEQ ID NO2078 GVPGGV SEQ ID NO1900 GGALGP SEQ ID NO1903 GLAGAG SEQ ID NO1906 AGLGAF SEQ ID NO1909 KAAKAG SEQ ID NO1912 RFPGVG SEQ ID NO1915 PQPGVP SEQ ID NO1918 GYGPGG SEQ ID NO1921 VLPGVG SEQ ID NO1924 AIPGIG SEQ ID NO1927 AAAAAA SEQ ID NO1930 GVPGVG SEQ ID NO1933 GAAVPG SEQ ID NO1936 VGGIPT SEQ ID NO1939 GVGTPA SEQ ID NO1942 GVAPGV SEQ ID NO1945 PGGVAA SEQ ID NO1948 PGVVGV SEQ ID NO1951 YGAAVP SEQ ID NO1954 GVGIPG SEQ ID NO1957 FGLVGA SEQ ID NO1960 LGGVLG SEQ ID NO1963 GGVAAR SEQ ID NO1966 LPGVYP SEQ ID NO1969 VGISPE SEQ ID NO1972 YPGGVL SEQ ID NO1975 VPGAGV SEQ ID NO1978 VFYPGA SEQ ID NO1981 PGGKPL SEQ ID NO1984 GAGLGA SEQ ID NO1987 AVTFPG SEQ ID NO1990 PGVGLP SEQ ID NO1993 PKAPGV SEQ ID NO1493 PKLPGG SEQ ID NO1998 TGVGPQ SEQ ID NO2001 GVPGVP SEQ ID NO2004 GIAGVG SEQ ID NO2007 VPGAGI SEQ ID NO2O1O YGARPG SEQ ID NO2013 VGVGGI SEQ ID NO2016 LVPGVG SEQ ID NO2019 VGLAPG SEQ ID NO2022 LVPGGP SEQ ID NO2025 VGVPGL SEQ ID NO2028 VLGGLG SEQ ID NO2031 VVGAGP SEQ ID NO2034 LGGLGV SEQ ID NO2037 AFQFPL SEQ ID NO2040 RPGFGL SEQ ID NO2043 FGPGVV SEQ ID NO2046 FPGALV SEQ ID NO2049 ALVPGG SEQ ID NO2052 VGAGVP SEQ ID NO2055 GGLGAL SEQ ID NO2058 VGAGPA SEQ ID NO2061 LGVGGL SEQ ID NO2064 FPLGGV SEQ ID NO2067 PGFGLS SEQ ID NO2070 GPGVVG SEQ ID NO2073 VGTPAA SEQ ID NO2076 or with any of the following sequences at the C-terminal of a peptide:

TABLE 26 C-terminal sequences of protease generated peptide fragments of Elastin. Elastin PGGVPG SEQ ID NO2079 GALGGG SEQ ID NO2081 GLGAFP SEQ ID NO2083 VPGGVA SEQ ID NO1974 RFPGVG SEQ ID NO1915 PKAPGV SEQ ID NO1493 LPYGYG SEQ ID NO1920 GIAGVG SEQ ID NO2007 AAAAKA SEQ ID NO2091 GVGVPG SEQ ID NO2092 VGGIPT SEQ ID NO1939 GVGVGG SEQ ID NO2097 GFPGFG SEQ ID NO2099 PGVGIS SEQ ID NO21O1 PGVGVA SEQ ID NO2075 GIGPGG SEQ ID NO2105 RAAAGL SEQ ID NO2108 GVPGLG SEQ ID NO2110 VLGGLG SEQ ID NO2031 PAAAAA SEQ ID NO1929 GLGVPG SEQ ID NO1961 RPGFGL SEQ ID NO2043 LSPIFP SEQ ID NO2119 GPGGVA SEQ ID NO2122 GLGALG SEQ ID NO2123 GALGPG SEQ ID NO2125 AFPAVT SEQ ID NO1989 AAAAYK SEQ ID NO2128 PGVPTG SEQ ID NO2131 PIKAPK SEQ ID NO2134 VGTPAA SEQ ID NO2076 GIGGIA SEQ ID NO2137 IPVVPG SEQ ID NO2139 GAGIPG SEQ ID NO1947 PTYGVG SEQ ID NO2142 IPTYGV SEQ ID NO2145 AGGFPG SEQ ID NO2147 VGVAPG SEQ ID NO1943 PGVGLA SEQ ID NO2020 GVAPGV SEQ ID NO1945 GLGVGG SEQ ID NO2153 GLGVGA SEQ ID NO2155 VGAGPA SEQ ID NO2061 GGLGVG SEQ ID NO2158 AGQFPL SEQ ID NO2160 GFGLSP SEQ ID NO2161 AAKFGA SEQ ID NO2164 AGLGAL SEQ ID NO2167 LKPVPG SEQ ID NO1905 ALVPGG SEQ ID NO2052 VLPGAR SEQ ID NO2172 PTGAGV SEQ ID NO2175 AGAAGK SEQ ID NO2176 PGAGLG SEQ ID NO2080 LGGGAL SEQ ID NO1902 LGAFPA SEQ ID NO1988 ADAAAA SEQ ID NO2084 VGVLPG SEQ ID NO2086 GVGPFG SEQ ID NO2088 AAAAAK SEQ ID NO2090 AIPGIG SEQ ID NO1927 VVGVPG SEQ ID NO2063 PVVPGA SEQ ID NO2093 GGIPTY SEQ ID NO2095 GVGGIP SEQ ID NO2098 FGVGVG SEQ ID NO2100 SPEAQA SEQ ID NO2102 GVGVAP SEQ ID NO2103 APGIGP SEQ ID NO2106 AAAGLG SEQ ID NO2029 GVPGFG SEQ ID NO2111 VGIPGG SEQ ID NO2032 VPGVGG SEQ ID NO2114 PGVGGL SEQ ID NO2116 GVAARP SEQ ID NO2041 AQAAAA SEQ ID NO2120 TPAAAA SEQ ID NO1928 LGALGG SEQ ID NO2056 VAPVGV SEQ ID NO2126 AVTFPG SEQ ID NO1990 AAKAGA SEQ ID NO2129 GVPTGA SEQ ID NO2132 KLPGGY SEQ ID NO2135 VPGVPG SEQ ID NO2057 GTPAAA SEQ ID NO2138 VGVPGA SEQ ID NO1931 PEAAAK SEQ ID NO2141 TYGVGA SEQ ID NO2143 PGAIPG SEQ ID NO2146 GIPGVA SEQ ID NO2148 VGLAPG SEQ ID NO2022 LAPGVG SEQ ID NO2078 GGVAAA SEQ ID NO2151 PGLGVG SEQ ID NO2154 ALAAAK SEQ ID NO2156 AGPAAA SEQ ID NO1958 LGVGGL SEQ ID NO2064 GGVAAR SEQ ID NO1966 PIFPGG SEQ ID NO2162 AAKYGA SEQ ID NO2165 GAGVPG SEQ ID NO2002 PGVGVG SEQ ID NO2169 RPGVGV SEQ ID NO1938 GGFPGF SEQ ID NO2173 VGGVPG SEQ ID NO1941 GAGLGA SEQ ID NO1987 GGGALG SEQ ID NO2082 LGAGLG SEQ ID NO1908 PGVLGG SEQ ID NO2085 VPTGAG SEQ ID NO2087 VPLGYP SEQ ID NO2089 IPGIGG SEQ ID NO2005 IGGIAG SEQ ID NO2006 GVPGVG SEQ ID NO1933 VVSPEA SEQ ID NO2094 GIPTYG SEQ ID NO2096 VGVPGL SEQ ID NO2028 GVGAGG SEQ ID NO2014 VAPGVG SEQ ID NO1944 APGVGL SEQ ID NO2104 VAPGIG SEQ ID NO2107 AAGLGA SEQ ID NO2109 AGVPGL SEQ ID NO2112 GAGPAA SEQ ID NO2113 GLGGLG SEQ ID NO2115 PAAAAK SEQ ID NO2117 AARPGF SEQ ID NO2118 GPGIPG SEQ ID NO2121 PGGVAA SEQ ID NO1948 ALGPGG SEQ ID NO2124 KPVPGG SEQ ID NO2127 VTFPGA SEQ ID NO2065 VPQPGA SEQ ID NO2130 AGVKPK SEQ ID NO2133 YGYGPG SEQ ID NO2136 GVGTPA SEQ ID NO1942 AAAAAA SEQ ID NO1930 GVPGAG SEQ ID NO2140 ARPGVG SEQ ID NO1937 YGVGAG SEQ ID NO2144 VGAGGF SEQ ID NO2015 GISPEA SEQ ID NO2149 VPGAPG SEQ ID NO2150 APGVGV SEQ ID NO1973 PGIGPG SEQ ID NO2152 LGVGVG SEQ ID NO2027 LGGLGA SEQ ID NO2157 GPAAAA SEQ ID NO1959 GVGGLG SEQ ID NO2159 VAARPG SEQ ID NO2042 IFPGGA SEQ ID NO2163 AAKAAK SEQ ID NO2166 GIPGGV SEQ ID NO2168 IPPAAA SEQ ID NO2170 ARPGFG SEQ ID NO2171 GLSPIF SEQ ID NO2174 GIPVVP SEQ ID NO1934

Vimentin

Several candidate proteases may be responsible for the digestion of vimentin in fibrotic tissue We have through a range of in vitro cleavages of pure native proteins determined that the enzymes listed in the following table cleaved vimentin at least at the cleavage sites at each end of the following sequences or at the cleavage sites marked ‘.’ or where no ‘.’ is shown, at the ends of the sequences:

TABLE 27 Vimentin fragments generated by specific proteases. Aminoacid residue Protease Sequence between cleavage sites numbers* MMP2, RLRSSVPGVR. SEQ ID NO2177 69-78 MMP8, Trypsin MMP2, RLRSSVPGVL. SEQ ID NO2178 69-78 MMP8, Trypsin MMP2, .LLQDSVDFSL SEQ ID NO2179 79-89 MMP8, Trypsin MMP2, .FADLSEAANR SEQ ID NO2180 295-304 MMP8, Trypsin MMP2 .ISLPLPTFSS SEQ ID NO2181 410-420 *in the human vimentin sequence

Accordingly, in a method of the invention, said peptide fragments preferably comprise a neo-epitope formed by cleavage of vimentin by a protease at an N- or C-terminal site, or where indicated a site marked by the sign in any one of the partial sequences of vimentin in Table 24.

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of vimentin.

Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:

TABLE 28 N-terminal sequences of protease generated peptide fragments of vimentin. Vimentin LLQDSV SEQ ID NO2182 FADLSE SEQ ID NO2183 ISLPLP SEQ ID NO2184 or with any of the following sequences at the C-terminal of a peptide:

TABLE 29 C-terminal sequences of protease generated peptide fragments of vimentin. Vimentin SVPGVR SEQ ID NO2185 SVPGVL SEQ ID NO2186

Further cleavage sites defining neo-epitopes that may be assayed in a similar manner can be identified by exposing collagens, elastin, CRP and proteoglycans or other fibrotic tissue proteins to any of the enzymes described herein and isolating and sequencing peptides thereby produced. Furthermore, assays may be based on the neo-epitopes generated adjacent the illustrated cleavage sites, i.e. in the C-terminal sequences that lead up to the N-terminal epitopes given above and the N-terminal sequences that connect to the C-terminal epitopes described.

Assays for more than one of the peptides described above may be conducted separately and their results combined or more than one of the peptides described above may be measured together.

The result of an assay according to the invention may be combined with one or more other measured biomarkers to form a composite index of diagnostic or prognostic value.

Generally, all previously known immunoassay formats can be used in accordance with this invention including heterogeneous and homogeneous formats, sandwich assays, competition assays, enzyme linked assays, radio-immune assays and the like. Thus, optionally, said method is conducted as a competition immunoassay in which said immunological binding partner and a competition agent are incubated in the presence of said sample and the competition agent competes with the peptide fragments in the sample to bind to the immunological binding partner.

Said competition agent may be (1) a synthetic peptide derived from the sequence of collagen type I, III, IV, V, or VI, or from CRP, or from any of the proteoglycans versican, lumican, perlecan, decorin and biglycan peptide, or a competition agent derived from (2) a purified native collagen type I, III, IV, V, or VI, or CRP, or any of the proteoglycans neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, perlecan, decorin and biglycan cleaved by proteases to reveal said neo-epitope.

One suitable method could be a competition immunoassay using monoclonal antibodies or antibody binding fragments binding to neo-epitopes of collagen type I, III, IV, V, VI, CRP, vimentin, or any of the proteoglycans neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments or neo-epitopes on peptide fragments from other proteins derived from fibrotic tissue. Appropriately selected synthetic peptides coated onto the solid surface of a microtitre plate could compete with the sample for binding to the monoclonal antibodies or binding fragments. Alternatively, purified, native collagen type I, III, IV, V, VI, CRP, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments carrying the neo-epitope recognised by the monoclonal antibody or binding fragment could be used on the solid surface. Yet another alternative is to immobilise the monoclonal antibody or binding fragment on the solid surface and then co-incubate the sample with a synthetic peptide appropriately linked to a signal molecule, e.g. horseradish peroxidase or biotin.

The sample may be a sample of serum, blood, plasma or other, e.g. fibrotic tissue biopsy.

Assays may be conducted as sandwich assays using a first immunological binding partner specifically reactive with a said neo-epitope and a second immunological binding partner reactive with the relevant protein to which the neo-epitope belongs. Optionally, said second immunological binding partner is directed to a second neo-epitope of the same protein.

In certain preferred methods the method further comprises comparing the determined level of said binding of said peptide fragments with values characteristic of (a) comparable healthy individuals and/or (b) a pathological fibrotic condition and optionally associating a higher level of the measured peptide (normally indicated by a higher level of binding) with a more severe degree of a said condition.

An aspect of the present invention relates to the development of monoclonal antibodies recognising neo-epitopes as described above, especially for collagen types I and IV. This can be achieved by immunising mice with synthetic peptides originating from the amino acid sequence of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan molecules (including the sequences listed above or sequences terminating therein), fusing the spleen-cells from selected mice to myeloma cells, and testing the monoclonal antibodies for binding to neo-epitopes on relevant synthetic peptides. Specificity for neo-epitopes can be ensured by requiring reactivity with a synthetic peptide and a lack of reactivity with either a C-prolongated form of the immunising peptide (for a C-terminal neo-epitope) or an N-terminal prolongated form of the immunising peptide (for an N-terminal neo-epitope). Antibodies for neo-epitopes may also be evaluated to establish a lack of binding capacity to native collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, pelecan and biglycan. Alternatively, specificity for a neo-epitope can be ensured by requiring the reactivity of the antibody to be negatively dependent on the presence of biotin or other functional groups covalently linked to one of the terminal amino acids.

The invention includes an immunological binding partner which is specifically immunoreactive with a neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan by a protease at a end-site in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above, and may be for instance a monoclonal antibody or a binding fragment thereof.

The invention includes a cell line producing a monoclonal antibody against a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan at the end-sites of sequences in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above.

The invention further provides a peptide comprising a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan in any one of the partial sequences of these proteins set out above. Such a peptide may be conjugated as a hapten to a carrier for producing an immune response to said peptide, or immobilised to a solid surface or conjugated to a detectable marker for use in an immunoassay.

The invention further comprises an isolated nucleic acid molecule coding for a peptide comprising a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above.

The invention further comprises a vector comprising a nucleic acid sequence comprising an expression signal and a coding sequence which codes for the expression of a peptide comprising a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, versican, lumican, decorin, perlecan and biglycan in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above and further includes a host cell transformed with such a vector and expressing a said peptide.

Yet another aspect of the invention relates to kits, which can be used conveniently for carrying out the methods described above. Such kits may include (1) a microtitre plate coated with synthetic peptide; (2) a monoclonal antibody or antibody binding fragment of the invention reactive with said synthetic peptide; and (3) a labelled anti-mouse IgG immunoglobulin. Alternatively, such kits may include (1) a microtitre plate coated with purified native collagen type I, III, IV, V, VI, CRP, vimentin, versican, lumican, decorin, perlecan and biglycan fragments; (2) a monoclonal antibody recognising a neo-epitope on collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments and reactive with said purified collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan, and biglycan fragments; and (3) a labelled anti-mouse IgG immunoglobulin. Alternatively, such kits may include (1) a microtitre plate coated with streptavidin; (2) a synthetic peptide linked to biotin; (3) a monoclonal antibody recognising a neo-epitope on collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments and reactive with said synthetic peptide; and (4) a labelled anti-mouse IgG immunoglobulin. Yet another alternative could be kits including (1) a microtitre plate coated with streptavidin; (2) a synthetic peptide linked to biotin; (3) a monoclonal antibody recognising a neo-epitope on collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments (and reactive with said synthetic peptide) and conjugated to horseradish peroxidase.

Thus, the invention includes an immunoassay kit comprising an immunological binding partner as described herein, especially in respect of collagens types I and IV, and a competition agent which binds said immunological binding partner, and optionally one or more of a wash reagent, a buffer, a stopping reagent, an enzyme label, an enzyme label substrate, calibration standards, an anti-mouse antibody and instructions.

The assays described herein are useful in the diagnosis of fibrosis in patients. In addition, the tests are useful for the assessment of disease progression, and the monitoring of response to therapy. The immunological binding partners of the invention may also be used in immunostaining to show the presence or location of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan cleavage products.

EXEMPLIFICATION Example 1 Collagen Type III Degraded with MMP-9 Method

Cleavage: Collagen type III isolated from human placenta was dissolved in 10 mM acetic acid (1 mg/ml). The protein solution was then passed through a filter (Microcon Ultracel YM-10) to remove fragment contaminations. MMP-9 was preactivated with 4-aminophenylmercuric acetate (APMA, Sigma) at 37° C. for 3 hours. After activations, collagen type III and MMP-9 were mixed 100:1 and incubated shaking for 3 days at 37° C.

The solution was analyzed by liquid chromatography/mass spectrometry (LC/MS) and the fragments were identified by performing Mascot Search. The peptide sequences were selected by homology search, ensuring no cross-reactivity to other or related proteins, as well as interspecies cross-reactivity.

Antibody design: The peptide sequences were synthesized and conjugated to ovalbumin (OVA). Mice were immunized ever 2-3 weeks, up to five. Antibody titers were checked by screening peptides, both selection and de-selection. When sufficient antibody titers were achieved, positive mice were selected for fusion, euthanized, and the spleen was disintegrated and B-cells were removed for fusion with myeloma cells. Selections of antibody producing cells were done by culturing and re-seeding the surviving chimera cells in single cell clones. Clones are selected by selection and de-selection peptides followed by native reactivity testing (FIG. 1), as neoepitopes are generated by synthetic small peptide sequences, which may not reflect the native proteins. An IgG subtype clone is selected for antibody production. Antibody purification is done by protein-G column.

Assay development: Optimal antibody concentrations are determined by checker-board analysis, with dilutions of antibody coating and screening peptide, in competitions ELISA. The different determination for the collagen degraded by MMP-9 (CO3) assay is shown in Table 30.

TABLE 30 Limit of Detection, Avarage Inter- and Intraassay variation of the CO3 assay. Limit of Detection 0.5 ng/ml Average Interassay variation 3.71% Average Intraassay variation 5.48%

Example 2 CO3 in Biological Relevant Samples CO3 Levels in Bile Duct Ligated Rats Compared to Sham Operated Rats.

Method: Forty female Sprague-Dawley rats (6 months old) were housed at the animal research facilities at Nordic Bioscience. The experiments were approved by the Experimental Animal Committee of the Danish Ministry of Justice, and were performed according to the European Standard for Good Clinical Practice (2008/561-1450). The rats were housed in standard type III-H cages at 18-22° C. with bedding and nest material (Altromin 1324; Altromin, Lage, Germany) and purified water (Milli-Q system; Millipore, Glostrup, Denmark) ad libitum. Rats were kept under conditions of a 12-hour light/dark cycle.

Liver fibrosis was induced by common BDL. In short: The rat was anaesthetized, the bile duct found, two ligations were performed around the bile duct followed by dissection between the ligations, the abdomen was closed. In sham operated rats, the abdomen was closed without bile duct ligation.

The rats were divided into 2 groups: Group 1 (10 BDL and 10 sham operated rats) were sacrificed after 2 weeks, and Group 2 (9 BDL and 10 sham operated rats) were sacrificed after 4 weeks. On completion of the study period (2, or 4 weeks), after at least 14 hours fasting, all surviving animals were asphyxiated by CO₂ and sacrificed by exsanguinations.

Blood samples were taken from the retro-orbital sinus of at least 14 hours fasting rats under light CO₂/O₂ anaesthesia at baseline and at termination. The blood were collected and left 30 minutes at room temperature to cloth, followed by centrifugation at 1500 g for 10 minutes. All clot-free liquid were transferred to new tubes and centrifuged again at 1500 g for 10 minutes. The serum were then transferred to clean tubes and stored at −80° C.

CO3 were measured in ×5 diluted serum samples from the rats. Sham and BDL levels were compared by Mann-Whitneys two-tailed nonparametric test (α=0.05) of statistical significance assuming normal distribution. CO3 levels increased significantly in the BDL groups compared to the Sham-operated animals. The results are shown in FIG. 2 a and b.

Example 3 CO3 in Different Fibrotic Diseases Human Serum

CO3 levels were measured in serum from human with three different fibrotic diseases: Chronic obstructed pulmonary disease (COPD), Scleroderma, and Hepatitis virus C(HCV). The serum samples were retrieved from Sera Laboratories International Ltd (SLI Ltd), UK.

CO3 levels were increased in the three different fibrotic diseases (FIG. 3)

Example 4 Antibody Development Detection of Marker CO3-610C

Type III collagen (Abcam, Cambridge, UK) was degraded in vitro by activated MMP-9 (Merck KGaA, Darmstadt, Germany) for 2 days. Degradation fragments were sequenced by LS-MS/MS and identified by MASCOT search. A specific peptide sequence ⁶¹⁰KNGETGPQ was selected for antibody production. The N-terminal of this sequence is residue 610 of human collagen type III. The synthetic peptide was conjugated to ovalbumin prior to subcutaneous immunization of 4-6 week old Balb/C mice with about 200 μL emulsified antigen and 50 μg CO3-610C (KNGETGPQGPGGC-OVA). Consecutive immunizations were performed at two week intervals until stable sera titer levels were reached in Freund's incomplete adjuvant. The mice were bled from the second immunization on. At each bleeding, the serum titer was measured and the mouse with highest anti-serum titer was selected for fusion. After the fourth immunization, this mouse was rested for one month and then boosted intravenously with 50 μg CO3-610C in 100 μL 0.9% sodium chloride solution three days before isolation of the spleen for cell fusion.

Monoclonal antibody producing clones were selected using a) immunogenic peptide: KNGETGPQGP-GGC-Ovalbumine (OVA) (807678), b) screening peptide KNGETGPQGP-PG-K-Biotin (807971), c) de-selection peptides KDGETGAAGPPGK-Biotin (118318) representing a type II collagen alpha 1 chain, KDGEAGAQGPPGK-Biotin representing a type I collagen alpha 1 chain degradation product, purchased from the Chinese Peptide Company, Beijing, China. The ELISA coat plate was obtained from NUNC (Thermofisher, Copenhagen, Denmark). Peptide conjugation reagents and buffers were produced by Pierce (Thermofisher, Copenhagen, Denmark).

Buffer used for dissolving the coating peptide was composed of the following: 40 mM Na

HPO

, 12 H

O, 7 mM KH

PO

, 137 mM NaCl, 2.7 mM KCl, 25 mM EDTA, 0.1% Tween 20, 1% BSA, 10% sorbitol, pH 7. For a serum assay, buffer containing the following chemicals was used: 8 mM Na

HPO

, 12 H

O, 1.5 mM KH

PO

, 13.7 mM NaCl, 2.7 mM KCl, 0.1% Tween 20, 1% BSA, 0.003% phenol red, pH 7.4. A different buffer used for a urine assay contained 400 mM TRIZMA, 0.05% Tween 20, 0.1% BSA, 0.36% Bronidox L5, pH 8.0. For both serum and urine assays we used a washing buffer composed of 25 mM TRIZMA, 50 mM NaCl, 0.036% Bronidox L5, 0.1% Tween 20, and reaction-stopping buffer composed of 0.1% H

SO

. ELISA-plates used for the assay development were Streptavidin-coated from Roche (Hvidovre, Denmark) cat.: 11940279. All ELISA plates were analyzed with the ELISA reader from Molecular Devices, SpectraMax M, (CA. USA).

In preliminary experiments, we optimized the reagents, their concentrations and the incubation periods by performing several checkerboard analyses. A 96-well ELISA plate coated with streptavidin was further coated with 5 ng/ml of the synthetic peptide KNGETGPQGP-Biotinylated dissolved in PBS-TBE buffer at 20° C. for 30 minutes by constant shaking at 300 rpm. After washing with washing buffer, 20 μl of sample was added, followed by 100 μl of peroxidase conjugated anti-human mAb-NB51-32 CO3-610C solution (23 pg/ml in incubation buffer). The plate was incubated for 1 hour at 20° C. during which time it was shaken at 300 rpm. This was followed by washing and finally, 100 μl tetramethylbenzinidine (TMB) (Kem-En-Tec cat.438OH) was dispensed and the plate incubated for 15 minutes in darkness and shaken at 300 rpm. In order to cease the reaction, 100 μl of stopping solution was added and the plate analyzed in the ELISA reader at 450 nm with 650 nm as reference.

A standard curve was performed by serial dilution of biotinylated-NB51-32 CO3-610C for a serum assay, and biotinylated-NB51-134 CO3-610C for a urine assay. Standard concentrations were 0, 0.33, 1, 3, 9, 27, 81 and 162 ng/ml.

We designate fragments detected using the immunoassays so obtained as CO3-610C as the amino acid K at the N-terminal of the sequence KNGETGPQGP is amino acid 610 of the human collagen III sequence.

Example 5 Comparison of CO3-610C and Other Biomarkers in Induced Liver Fibrosis in Rats Animals

40 female Sprague-Dawley rats aged 6 months were housed at the animal research facilities at Nordic Bioscience, Copenhagen, Denmark. The experiments were approved by the Experimental Animal Committee of the Danish Ministry of Justice and were performed according to the European Standard for Good Clinical Practice (2008/561-1450). The rats were housed in standard type III-H cages at 18-22° C. with bedding and nest material (Altromin 1324; Altromin, Lage, Germany) and water ad libitum. Rats were kept under conditions of a 12-hour light/dark cycle.

Study Design

In 20 rats, liver fibrosis was induced by common BDL. The surgical procedure was performed under sterile conditions. The rat was anaesthetized, the bile duct localized and ligated in two places followed by dissection between the ligations, and the abdomen was closed. The other 20 rats were subjected to a sham operation, in which the abdomen was closed without bile duct ligation. The rats were then divided into 2 groups: Group 1 (10 BDL rats and 10 sham-operated rats) was sacrificed after 2 weeks and Group 2 (10 BDL and 10 sham-operated rats) was sacrificed after 4 weeks. On completion of the study period (2 or 4 weeks), after at least 14 hours fasting, all surviving animals were asphyxiated by CO₂ and sacrificed by exsanguinations.

Blood Sampling

Blood samples were taken from the retro-orbital sinus of rats after at least 14 hours fasting, under light CO₂/O₂ anaesthesia, at baseline and at termination. Blood was left 30 minutes at room temperature to clot, followed by centrifugation at 1500 g for 10 minutes. All clot-free liquid was transferred to fresh tubes and centrifuged again at 1500 g for 10 minutes. The serum was then transferred to clean tubes and stored at −80° C.

Tissue Handling

After the rats were put down, their livers were carefully dissected, weighed, fixed in 4% formaldehyde for a minimum of 24 hours, cut into appropriate slices and embedded in paraffin. Sections 5 μm thick were cut, mounted on glass slides and stained with Sirius Red. The liver sections were evaluated histologically by assessment of the architecture, presence of inflammation, proliferation of bile ducts and fibrosis. The de novo bile duct formation in the parenchyma was evaluated semi-quantitatively using the following scoring system: normal=0, mild changes (⅓ or less of the lobule affected)=1, moderate changes (between ⅓ and ⅔ of the lobule affected)=2, and severe changes (⅔ or more of the lobule affected)=3. Digital photographs were captured using an Olympus BX60 microscope with ×40 and ×100 magnification and an Olympus 5050-zoom digital camera (Olympus, Tokyo, Japan).

Determination of Total Collagen and Serum CTX-II

The total collagen concentration was assayed using the commercial QuickZyme Collagen Assay (QuickZyme Bioscience, Leiden, The Netherlands). The concentration of CTX-II was assayed using the commercial Rat CTX-II kit (IDS Nordic, Herlev, Denmark). All samples were assayed in duplicate.

Type III Collagen mRNA Quantification

The number of transcripts of type III collagen (Col3a1) in liver tissue samples was determined by quantitative real-time polymerase chain reaction (RT-PCR) using fluorescent reporter probes. The number of Col3a1 copies in the sample was extrapolated from a standard curve obtained using Col3a1 plasmid cDNA Image Clone 7097081 (Geneservice, Cambridge, UK) as dilution standard. Amounts of Col3a1 were normalized with those of housekeeping gene hypoxanthine phosphoribosyltransferase 1 (Hprt1). Primers and probes for Col3a1 and Hprt1 mRNAs were designed using NCBI Reference Sequences NM_(—)032085.1 and NM_(—)012583.2 as templates, respectively (TIB Molbiol GmbH, Berlin, Germany). Total RNA was extracted from frozen liver samples using Absolutely RNA Miniprep kit (Stratagene, La Jolla, Calif., USA) following the manufacturer's instructions and its quality assessed in RNA Nano chips using a 2100 Bioanalyzer instrument (Agilent Technologies, Santa Clara, Calif., USA). Immediately after RNA isolation, complementary DNA (cDNA) was synthesised with Transcriptor First Strand cDNA Synthesis kit (Roche, Basel, Switzerland) using 1 μg of RNA as the template. For each sample tested, a cDNA synthesis negative control, omitting reverse transcriptase enzyme from the reaction mix, was included. Separate PCR reactions for Col3a1 and Hprt1 were performed in a 20 μL format using the Lightcycler Faststart DNA Master Plus Hybprobe kit (Roche) according to the manufacturer's instructions. Real time fluorescence data was collected in a Lightcycler 2.0 instrument (Roche).

Extractions

Tissue was pulverized in excess of liquid nitrogen in a steel mortar. Samples were then transferred into a 1.5 ml eppendorf tube and left shaking overnight at 4° C. in 0.5M Acetic Acid solution containing protease inhibitor cocktail (Roche). The samples were then sonicated with ultrasounds using 5 pulses at 60% amplitude (U50 control, IKA Labortechnik) and left for an additional 2 hours at 4° C. after which they were centrifuged for 5 minutes at 13,000 rpm. Supernatant was carefully removed, transferred in a new eppendorf and stored at −80° C.

Densitometry

Densitometry measurements were performed using UN-SCAN-IT Version 6.1 from Silk Scientific (give city, country).

Histology Image Analysis

Histology sections stained with Sirius Red were analyzed using Visiopharm software Version 3.2.8.0 (give city, country). Images were acquired using Pixelink PL-A623C microscope digital camera.

SDS PAGE and Western Blots

20 μg of tissue extract was mixed with loading buffer (Invitrogen LDS 4×, NP0007) containing the reducing agent (NuPAGE, NP0004 from Invitrogen). Samples were then loaded into 4-12% Bis-Tris gradient gel (NP0332BOX from Invitrogen) and run for 52 minutes at 200V. Proteins were then transferred onto a nitrocellulose membrane using the i-Blot transfer system (Invitrogen), blocked with 5% milk in (? Need to spell out?) TTBS overnight at 4 degrees. Beta Actin antibody (AbCam ab8229, give company, city country?) was used as a loading control.

Statistical Analysis

Mean values and standard error of the mean (SEM) were calculated using GraphPad Prism (GraphPad Software, Inc., San Diego, Calif., USA) and compared by Student's two-tailed paired t-test (α=0.05) of statistical significance assuming normal distribution, or by Mann-Whitney two-tailed non-parametric test (α=0.05). The coefficient of correlation (R²) and the corresponding p-value was determined by linear regression.

Results

Liver appearance: At the time of sacrifice, livers of control animals showed normal gross morphology while livers of BDL animals were enlarged. The liver weights were significantly increased in BDL rats compared to the sham-operated controls (mean weights at sacrifice: 2 weeks post-surgery, sham 8.1 g; BDL 14.1 g; 4 weeks post-surgery, sham 9.0 g; BDL 19.4 g) (FIG. 4 panel A). Semi-quantitative scoring of liver sections using the 0-3 scale showed significantly more structural changes of the liver at 4 weeks compared with 2 weeks (FIG. 4 panel B). FIG. 4, panel A shows liver weight in bile duct ligation (BDL)- or sham-operated rats. Data are shown as mean±SEM. [*** , P<0.0001. Panel B shows scoring of the structural changes in the liver of each group. Data are shown as mean±SEM. **, P=0.0094. Panel C shows Sirius Red photomicrographs showing the hepatic structure in sham-operated rats, and in BDL rats 2 and 4 weeks post-surgery. The hepatic structure around the portal tract is clearly disrupted in BDL rats compared with the sham-operated rats. Collagens are highlighted in red. Original magnification was ×40.

Under histological examination, the livers of sham-operated animals showed no sign of fibrosis and were microscopically normal (FIG. 4C). In the BDL livers, a marked ductal proliferation was observed. In the 2-week post-surgery group, the proliferation was located around the portal tract while in the 4-week group the proliferation had spread (FIG. 4C). Collagen deposition was found around the ductular structures. Inflammation was minimal and confined to the portal tracts. No signs of cholestasis were seen, whether intracellular cholestasis, bile plugs, bile infarctions or hepatocytic rosette formation.

Changes in CO3-610C levels: FIG. 5 shows in panel A MMP-9 mediated CO3 degradation serum levels in bile duct ligated (BDL)- or sham-operated rats. Data are shown as mean±standard error of mean. 2 weeks post-surgery ***P<0.0001 and 4 weeks post-surgery **P=0.0014. In panel B are shown CO3-610C delta values (termination-baseline paired), 2 weeks post-surgery P<0.0001 and 4 weeks post-surgery P=0.0016. In panel C are shown CTX-II levels in BDL- or sham-operated rats. Data are shown as mean±standard error of mean.

In the BDL groups CO3-610C levels increased significantly compared to sham groups (mean values: 2 weeks, post-surgery sham 39.7 ng/ml, BDL 100.3 ng/ml; average increase between the groups was 153%; 4 weeks post-surgery, sham 39.7, BDL 92.6 ng/ml, average increase between the groups was 133%) (FIG. 5 panels A and B). There were no changes in the sham groups. CTX-II levels indicating collagen type II degradation did not change in the sham or BDL groups (FIG. 5 panel C).

Type III Collagen Gene Expression: FIG. 6 shows Type III collagen gene expression in BDL or sham-operated rats. Data are shown as mean±standard of mean; 2 weeks post-surgery P<0.0001 and 4 weeks post-surgery P=0.0006

Type III collagen a1 chain mRNA increased significantly in both BDL groups compared with sham-operated rats.

Western Blot and Densitometry: FIG. 7 shows changes in the expression of CO3-610C in the liver of rats in BDL- and sham-operated groups assessed by A) Western blot 2 and 4 weeks post-surgery and B) Bands from western blot quantified by densitometry.

Western blot analysis showed very low levels of CO3-610C in sham-operated rats (FIG. 7 panel A). At and after 2 weeks post-surgery CO3-610C levels prominently increased (FIG. 7 panel A). Results were quantified by densitometry analysis (FIG. 7 panel B).

Histology image analysis: FIG. 8 panel A shows in the top row histology sections from BDL- or sham-operated rats stained with Sirius Red. The bottom row shows masked histology sections for quantifying total collagen content (red colour) in the liver. Panel B shows total collagen quantified by Visiopharm software—2 weeks post-surgery P=0.0081; 4 weeks post-surgery P=0.0047

Histology sections stained with Sirius Red and enhanced using Visiopharm software showed increasing collagen content over time in BDL-operated rats.(FIG. 8 panel A). The red color in the mask representing collagen was quantified using the same software (FIG. 8 panel B) and confirmed a significant increase in total collagen content in BDL-operated rats compared with sham-operated rats (2 weeks post-surgery P=0.0081; 4 weeks post-surgery P=0.0047).

Correlation: FIG. 9 panel A shows a correlation of Col3a1 to CO3-610C was found with R²=0.6993, P<0.0001. In panel B, a correlation of CO3-610C to % collagen was found with R²=0.2278 and P=0.0050. In panel C a correlation of Col3a1 to % collagen was found with R²=0.5409, P<0.0001.

Correlations were found of the following: Col3a1 mRNA to CO3-610C with R²=0.6993 and P<0.0001 (FIG. 9A), and CO3-610C to % collagen quantified by visiopharm with R²=0.2278 and P=0.0050 (FIG. 9B), and Col3a1 mRNA to % collagen quantified by visiopharm with R²=0.5409 and P<0.0001 (FIG. 9C).

ECM remodelling is an integrated process of tissue development, maintenance and pathogenesis. Proteolytic activity is essential in this process for cell migration, removal of damaged tissue, and sequestering of new proteins, for the correct and optimal tissue orientation and quality (108:109). The specific matrix degradation products, neo-epitopes, may be important for the identification of new biochemical markers of liver fibrosis matrix turnover and understanding fibrosis pathogenesis. At present there are no available measuring techniques, nor biochemical markers, that allow for assessment of ECM remodeling in the pathogenesis of fibrosis.

In this example, to investigate the CO3-610C marker under in vivo situations, 6 months BDL rats were chosen, as they previously have been shown to have a lower collagen remodelling compared to younger rats. The rats are skeletally mature, and the growth plate is almost dormant, thereby contributing to a much lower extent to the overall collagen turnover. This influences the sensitivity and specificity for biomarker. These rats clearly presented with hepatic fibrosis, as evaluated by both quantitative histological analysis, and enlargement with increased weight, thus the model was an appropriate one to look for evidence of ECM remodeling, in particular for evidence of collagen type III in serum.

The present data clearly demonstrate the neo-epitope CO3-610C from MMP-9 mediated collagen type III degradation is a diagnostic biochemical marker for liver fibrosis with an average increases in serum of up to 153% from sham to BDL-operated rats.

To further investigate the biological rationale for the increased CO3-610C marker, we did protein extractions from healthy and diseased livers. By western blotting, we identified a predominant band, suggesting this to be an abundant protein fragment in diseased but not healthy livers. This provides evidence for the pathological accuracy of this novel marker.

To further investigate the pathological turnover representation of the liver, we measured type III collagen mRNA. We found an increase of mRNA in the BDL rats compared to those undergoing the sham operation, which correlates with previous findings. These data strongly suggest that liver fibrosis is not only an accumulation of ECM proteins, but also an accelerated turnover situation, in which both tissue formation and tissue degradation both are highly up regulated. Tissue formation outstrips tissue degradation, leading to an accumulation of scar tissue over time. Previous investigators have used other matrix turnover proteins to assess liver fibrosis, one being the type III collagen formation marker N-terminal type III pro-collagen. This marker represents collagen type III formation and has shown to be increased in liver fibrosis in previous studies.

To further understand and the dynamics of the biochemical makers CO3-610C, we did a range of correlations. Most importantly, there was a significant correlation of CO3-610C to the extent of fibrosis measured in the liver by quantitative histology. The level of liver fibrosis was correlated to the expression levels of the mRNA of collagen type III. Finally, the CO3-610C correlated to mRNA of collagen type III in the liver. Taken together, there was a significant correlation of the pathological processes in the liver with the levels of the systemic biochemical markers CO3-610C. In addition the tissue extractions provided evidence that the circulation levels were locally produced.

Example 6 ELISA on Human Serum Samples

Human serum samples were obtained from patients with Chronic Obstructive Pulmonary Disease (COPD) (n=5), scleroderma (n=5), chronic hepatitis C virus infection (n=5), and healthy controls (n=5). The serum samples were tested in the CO3-610 ELISA (see Example 4 above) to determine the concentration of CO3-610 fragments. Results are shown in FIG. 10. While serum samples from the healthy subjects had concentration of CO3-610 fragments below 30 ng/ml, the diseased subjects were found to have elevated levels in circulation suggesting massive tissue remodelling in the affected fibrotic tissues.

Example 7 Reactivity of Clone nb94

Mice were immunized with synthetic peptide KAFVFP (SEQ ID NO1167) conjugated to ovalbumin (KAFVFPKESD-GGC-OVA (SEQ ID NO1049)), spleen cells were used for fusion, and monoclonal antibodies tested for reactivity to biotinylated KAFVFP (SEQ ID NO 1167), i.e. (KAFVFPKESD-biotin (SEQ ID NO1049)) immobilized in wells of microtitre plates precoated with streptavidin. Antibodies binding to biotinylated KAFVFPKESD (SEQ ID NO1049), which could be inhibited by co-incubation with KAFVFPKESD (SEQ ID NO1049) but not the elongated peptide RKAFVFPKESD (SEQ ID NO1166), were selected for further characterization. The preferred monoclonal antibody was designated NB94-37-1A7. Using a competition ELISA, essentially as described above with biotinylated KAFVFPKESD (SEQ ID NO1049) (used at 0.15 ng/ml) immobilized in the wells of streptavidin-coated microtitre plates, an incubation step (90 minutes at 20° C.) with sample and monoclonal antibody NB94-37-1A7 followed by a washing step, and then addition of peroxidase-conjugated anti-mouse immunoglobulins. For competition the following material was used in 2-fold dilutions; (1) the synthetic KAFVFP (SEQ ID NO1167) peptide; (2) a nonsense peptide (KNEGTG) unrelated to CRP; (3) a pool of human serum samples; (4) CRP proteolytically cleaved with MMP3 for 7 days, subsequently stopped by addition of EDTA to block protease activity, and stored at −80° C. until testing; (5) same as (4) but using MMP8 instead of MMP3; (6) same as (4) except using Cathepsin K (for 2 days) instead of MMP3 (and E64 as inhibitor to block Cathepsin K activity).

The data demonstrate that monoclonal antibody NB94-37-1A7 binds strongly to the synthetic peptide KAFVFPKESD (SEQ ID NO1049), and with CPR cleaved with MMP3 and MMP8. Cleavage of CRP with Cathepsin K release less analyte recognized by monoclonal antibody NB94-37-1A7. Finally, the data shows that the antibody binds to peptide fragments in human serum confirming the presence of this sequence in circulating peptide fragments.

Example 8 CO3 in Biological Relevant Samples: CO3 Levels in Carbon Tetrachloride (CCl4)-Induced Cirrhosis in Rats Animals and Induction of Cirrhosis:

This study included 52 male Wistar rats with fibrosis or cirrhosis and 35 male Wistar control rats. To cause them to develop fibrosis or cirrhosis three-month old animals were included in an induction program with carbon tetrachloride (CCl4) and Phenobarbital treatment. CCl₄ was administered by inhalation twice weekly and phenobarbital (0.3 g/l) added to the drinking water. Animals were allowed free access to water and food throughout the study.

Fibrosis Quantification:

Liver sections (4 μm) were stained in 0.1% Sirius red F3B (Sigma-Aldrich, St. Louis, Mo.) in saturated picric acid (Sigma-Aldrich). Relative fibrosis area (expressed as a percentage of total liver area) was assessed by analyzing 36 fields of Sirius red-stained liver sections per animal. Each field was acquired at 10× magnification [E600 microscope (Nikon) and RT-Slider SPOT digital camera (Diagnostic Instruments, Inc., Sterling Heights, Mich.). Results were analyzed using a computerized Bioquant Life Science morphometry system. To evaluate the relative fibrosis area, the measured collagen area was divided by the net field area and then multiplied by 100. Subtraction of vascular luminal area from the total field area yielded the final calculation of the net fibrosis area. From each animal analyzed, the amount of fibrosis as percentage was measured and the average value presented.

Classification of Groups According to their Fibrosis/Cirrhosis Stage:

Animals were classified into 4 different stages of fibrosis and cirrhosis (Group A: moderate fibrosis, group B: advanced fibrosis, Group C: moderate cirrhosis, and Group D: advanced cirrhosis) that were determined by the percentage of Sirius red positive liver area (Group A: <5%, Group B: 5 to 10%, Group C: 10 to 15% and Group D: >15%). For this purpose, control and fibrotic/cirrhotic rats were studied considering four different time points during the CCl4 treatment: 8, 12, 16 and 20 weeks after starting the cirrhosis induction program.

Hyaluronic Acid Measurement:

Serum hyaluronan was measured using a sandwich ELISA kit (R&D Systems Inc., Minneapolis, Minn., USA).

Statistics:

Statistical analysis of results was performed by unpaired Student's t tests when appropriate. Data were expressed as mean±S.E.M., and they were considered significant at a p level of 0.05 or less.

Study Design:

Animals included in this protocol were randomly assigned to one of the following groups: A/ eight weeks of CCl₄ treatment, B/ twelve weeks of CCl₄ treatment, C/ sixteen weeks of CCl₄ treatment and D/ twenty weeks of CCl₄ treatment. In parallel, four control groups were studied at the same time points. Thirteen fibrotic rats and seven control rats were included in each group. At the end of the study, rats were placed in standard metabolic cages (Tecniplast Deutschland, Hohenpeissenberg, Germany) during an adaptation period of 3 days before proceeding with the twenty-four-hour urine collection. Urinary volumes were determined gravimetrically. During the adaptation period, rats were allowed to get free access to tap water and food. Then, 24-hour urine samples were centrifuged for 5 min at 2,500 rpm and aliquoted into ten polypropylene tubes (400 μL each). Urine samples were stored at −80° C. for subsequent analysis.

At scheduled necropsies, rats were weighed, anesthetized with pentobarbital (50 mg/kl) and decapitated. Blood were collected and allowed to stand at room temperature for 20 min to allow clotting and then centrifuged for 10 min at 2500 rpm. Serum were collected in polypropylene tubes aliquots (400 μl each) and transferred via dry ice to a −80° C. freezer. Collection of baseline blood samples at the beginning of the CCl₄ treatment was not considered in order to avoid additional intervention that may increase the risk of infection and/or introduce modifications in the experimental model that may compromise the evolution of the induced pathophysiological process. For histology and Sirius red staining, half of the left lobe of the liver were placed in 10% neutral buffered formalin for 16 hours, embedded in paraffin and sectioned into 4-μm-thick slices. After liver fibrosis quantification, the unused paraffin block material was preserved for biomarker quantification. The other half of the left lobe was flash-frozen in liquid nitrogen and stored for Western blot, RT-PCR or immunohistochemical analysis. Measurements of liver fibrotic area, serum and urine osmolality, Na⁺ and K⁺, albumin, creatinine, alanine amino-transferase and lactate dehydrogenase were made according to the Material and Methods section.

Results:

Histological Validation of the Model:

Liver collagen was quantified in all study animals by Sirius red staining of liver slices. The final data for each animal was taken as the average of red staining observed in 36 consecutive microscope fields (FIG. 12).

FIG. 12 shows representative pictures from two sets of 36 images used to quantify collagen accumulation in liver in rat #1 (left) and rat #43 (right) treated with carbon tetrachloride for eight and twenty weeks respectively.

The serum CO3 marker shows statistically significant increases in both fibrotic and cirrhotic rats compared to control rats. Animals were classified according to a fully automated syrius red staining of the liver procedure used to quantify fibrosis (FIGS. 13 and 14).

FIG. 13 shows serum CO3 levels in CCl₄ inhalation and control rats as performed in Hospital Clinic (Barcelona). Each point represents one animal. Rats were classified according a computerized image analysis method of syrius red staining of the liver used to quantify fibrosis.

When quantitative values of serum CO3 and syrius red staining of the liver were studied in each individual animal, we found a statistically significant correlation between the two variables (R2=0.4087; n=21) (FIG. 14).

We have compared the levels of CO3-610C with the serological benchmark of liver fibrosis hyaluronic acid (HA). HA levels were quantified with a commercial ELISA kit and results show significant elevations of this ECM component in cirrhotic rats vs. fibrotic animals (FIGS. 15 and 16).

The correlation of CO3 to Sirius red outperformed that of HA. More than seventy percent of the variation in liver fibrosis histological quantification can be explained by the serological measurement of CO3. The remaining thirty percent is due to unknown variables or inherent variability. Instead only 25% of liver fibrosis can be explained by measuring hyaluronic acid (FIG. 15).

As expected from the previous result no correlation could be found between CO3 and hyaluronic acid suggesting that they are the result of two independent pathophysiological processes in the development of liver fibrosis (FIG. 17).

Example 9 Bleomycin Induced Skin Fibrosis in Mice

Mice were treated by application to the skin of PBS or bleomycin. Increasing levels in urine of the MMP-9 mediated collagen III (CO3) degradation fragment CO3-610 were associated with skin fibrosis progression in mice.

FIG. 18 shows a skin section from a PBS treated mouse at 8 weeks of treatment (panel A) and a skin section from Bleomycin treated mouse at 8 weeks of treatment (panel B). Skin thickness increase between PBS (n=7/time point) and Bleomycin (n=13/time point) treated mice for 2 weeks (P=0.0029), 4 weeks (P=0.0004), 6 weeks (P<0.0001) and 8 weeks (P<0.0001) is plotted in panels C and D. Overall skin thickness increase between PBS (n=28) and Bleomycin (n=52) treated mice for the duration of the study (P<0.0001). Skin width was calculated by Visiopharm software as an overall number per skin section instead of sampling pictures.

FIG. 19 shows CO3-610 urine assay results which demonstrate a significant increase throughout the time points of the study. The figure shows result per time point (n=7 PBS, n=13 Bleomycin treated per termination point) and collective CO3-610 levels for all time points (n=28 PBS and n=52 Bleomycin treated mice). 2 weeks P=0.0008, 4 weeks P<0.0001, 6 weeks P<0.0001, 8 weeks P<0.0001 and overall P<0.0001.

FIG. 20 shows a CO3-610 Western blots image with control C and Bleomycin B after 2 and 8 weeks treatment (panel A). CO3-610 densitometry measurements for all time points (n=7 PBS and n=13 Bleomycin treated per termination point) and collective CO3-610 levels (n=28 PBS and n=52 Bleomycin treated mice) are shown in panel B, demonstrating a statistically significant increase of CO3-610 levels (P<0.0001).

As seen in FIG. 21, CO3-610 levels in urine assay were found to be correlated with skin thickness progression, and therefore total collagen deposition r=0.4883, R2=0.2384.

As seen in FIG. 22, statistically significant correlation was found (r=0.6528, P<0.0001) between results from the CO3-610 ELISA urine assay and Western blot densitometry measurements.

In this specification, unless expressly otherwise indicated, the word ‘or’ is used in the sense of an operator that returns a true value when either or both of the stated conditions is met, as opposed to the operator ‘exclusive or’ which requires that only one of the conditions is met. The word ‘comprising’ is used in the sense of ‘including’ rather than in to mean ‘consisting of’. All prior teachings acknowledged above are hereby incorporated by reference. No acknowledgement of any prior published document herein should be taken to be an admission or representation that the teaching thereof was common general knowledge in Australia or elsewhere at the date hereof.

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EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by this invention and the following claims.

INCORPORATION BY REFERENCE

All publications, patent applications and patents identified herein are expressly incorporated herein by reference in their entirety. 

1. A method of diagnosis or of quantitation of fibrosis comprising obtaining a patient biofluid sample, conducting an immunoassay to measure neo-epitope containing protein fragments naturally present in said sample, and associating an elevation of said measure in said patient above a normal level with the presence or extent of fibrosis, wherein said immunoassay is conducted by a method comprising: contacting protein fragments naturally present in said sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to said immunological binding partner to measure therein protein fragments comprising said neo-epitope, and wherein said protein is collagen type III, collagen type I, collagen type IV, collagen type V, or collagen type VI, elastin, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, vimentin, or C-reactive protein, subject to the proviso that when the neo-epitopes are formed by cleavage of type I collagen, the cleavage is not at a site at which collagen type I is cleaved by cathepsin K.
 2. A method as claimed in claim 1, wherein said immunological binding partner specifically binds fragments of collagen type III having an N-terminal sequence KNGETG . . . .
 3. A method as claimed in claim 1, wherein said immunological binding partner specifically binds fragments of CRP having an N-terminal sequence KAFVFP . . . (SEQ ID NO1167).
 4. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of collagen type III, said peptides comprising an N-terminal sequence selected from the group consisting of: Collagen type III GIPGAP . . . SEQ ID NO372 IAGITG . . . SEQ ID NO375 KGDAGQ . . . SEQ ID NO381 GKSGDR . . . SEQ ID NO383 DGTSGH . . . SEQ ID NO135 GPPGVA . . . SEQ ID NO158 GARGLA . . . SEQ ID NO111 KGESGK . . . SEQ ID NO374 QPGVMG . . . SEQ ID NO144 GPPGPT . . . SEQ ID NO391 GLPGPP . . . SEQ ID NO394 GINGSP . . . SEQ ID NO397 LMGARG . . . SEQ ID NO400 DKGEPG . . . SEQ ID NO403 PGMKGH . . . SEQ ID NO406 FPGMKG . . . SEQ ID NO409 GQPGDK . . . SEQ ID NO412 GERGSP . . . SEQ ID NO415 GSDGQP . . . SEQ ID NO405 GFPGAP . . . SEQ ID NO420 PGPQGH . . . SEQ ID NO426 PGPPGI . . . SEQ ID NO432 GPPGSN . . . SEQ ID NO423 PQGLQG . . . SEQ ID NO440 GAPGFR . . . SEQ ID NO435 GAPGPQ . . . SEQ ID NO445 GPTGPI . . . SEQ ID NO448 KGSPGA . . . SEQ ID NO444 GSRGAP . . . SEQ ID NO462 NTGAPG . . . SEQ ID NO437 HAGAQG . . . SEQ ID NO434 PGPQGP . . . SEQ ID NO453 AGQPGE . . . SEQ ID NO454 VKGERG . . . SEQ ID NO159 GPPGAP . . . SEQ ID NO461 GSPGAQ . . . SEQ ID NO464 PGAPGL . . . SEQ ID NO467 ESCPTG . . . SEQ ID NO433 GPAGIP . . . SEQ ID NO441 GDPGPP . . . SEQ ID NO373 IKGHRG . . . SEQ ID NO376 ITGARG . . . SEQ ID NO379 LQGLPG . . . SEQ ID NO384 IGSPGP . . . SEQ ID NO AGPPGM . . . SEQ ID NO145 GAPGEK . . . SEQ ID NO141 GLSGER . . . SEQ ID NO387 IPGAPG . . . SEQ ID NO117 INGSPG . . . SEQ ID NO392 KNGETG . . . SEQ ID NO395 PGENGK . . . SEQ ID NO398 GKDGES . . . SEQ ID NO418 GHAGAQ . . . SEQ ID NO404 FPGARG . . . SEQ ID NO407 PGDKGE . . . SEQ ID NO410 GPPGEN . . . SEQ ID NO413 PGVPGA . . . SEQ ID NO416 GPPGPP . . . SEQ ID NO100 GAAGEP . . . SEQ ID NO421 PGFPGM . . . SEQ ID NO427 GITGAR . . . SEQ ID NO430 RPGLPG . . . SEQ ID NO436 GPPGVA . . . SEQ ID NO158 PGFRGP . . . SEQ ID NO443 GFPGNP . . . SEQ ID NO446 GDAGQP . . . SEQ ID NO449 GSPGER . . . SEQ ID NO439 TGARGL . . . SEQ ID NO378 VGGLAG . . . SEQ ID NO155 PGAPGG . . . SEQ ID NO455 AGQQGA . . . SEQ ID NO457 GLAGPP . . . SEQ ID NO388 GGAGEP . . . SEQ ID NO463 SPGAQG . . . SEQ ID NO465 IKGPAG . . . SEQ ID NO169 GIPGQP . . . SEQ ID NO468 DAGAPG . . . SEQ ID NO469 LAGPPG . . . SEQ ID NO89 RGLAGP . . . SEQ ID NO377 VKGESG . . . SEQ ID NO380 LRGGAG . . . SEQ ID NO382 AIGSPG . . . SEQ ID NO143 LSGERG . . . SEQ ID NO176 PQGPPG . . . SEQ ID NO389 YQGPPG . . . SEQ ID NO4O1 FRGPAG . . . SEQ ID NO137 GPPGEP . . . SEQ ID NO393 LPGIAG . . . SEQ ID NO396 LKGENG . . . SEQ ID NO399 QQGAIG . . . SEQ ID NO390 GERGAP . . . SEQ ID NO402 GFPGAR . . . SEQ ID NO408 GDKGET . . . SEQ ID NO411 AAGFPG . . . SEQ ID NO417 GARGND . . . SEQ ID NO419 GGAGPP . . . SEQ ID NO425 GARGPP . . . SEQ ID NO422 GSPGGP . . . SEQ ID NO428 GIAGIT . . . SEQ ID NO431 GAPGPM . . . SEQ ID NO438 SGDRGE . . . SEQ ID NO429 GPVGPS . . . SEQ ID NO447 GPPGIN . . . SEQ ID NO470 NGEKGE . . . SEQ ID NO450 AIGPSG . . . SEQ ID NO368 ERGLPG . . . SEQ ID NO385 VAGPPG . . . SEQ ID NO452 GIPGFP . . . SEQ ID NO414 PGPPGP . . . SEQ ID NO458 GRNGEK . . . SEQ ID NO460 SPGGKG . . . SEQ ID NO459 PGVSGP . . . SEQ ID NO466 PGAPGQ . . . SEQ ID NO456 SRGAPG . . . SEQ ID NO451 GPKGDA . . . SEQ ID NO424


5. method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of collagen type III, said peptides comprising an C-terminal sequence selected from the group consisting of: Collagen type III . . . GPPGPA SEQ ID NO94 . . . GMPGPR SEQ ID NO473 . . . ERGAAG SEQ ID NO476 . . . ERGPPG SEQ ID NO147 . . . GLPGLA SEQ ID NO486 . . . GLAGTA SEQ ID NO488 . . . LAGPPG SEQ ID NO89 . . . IPGFPG SEQ ID NO492 . . . FPGPKG SEQ ID NO491 . . . GPPGIC SEQ ID NO2187 . . . PGPQGL SEQ ID NO497 . . . SPGPKG SEQ ID NO499 . . . LPGPPG SEQ ID NO72 . . . GHRGFD SEQ ID NO503 . . . GLPGIA SEQ ID NO507 . . . GLPGPP SEQ ID NO394 . . . TGARGL SEQ ID NO378 . . . PQGLPG SEQ ID NO508 . . . GTPGLQ SEQ ID NO521 . . . GMKGHR SEQ ID NO531 . . . EMGPAG SEQ ID NO534 . . . GVKGER SEQ ID NO538 . . . GPPGPR SEQ ID NO544 AGPRGA SEQ ID NO547 . . . GRNGDP SEQ ID NO171 . . . AGIPGF SEQ ID NO496 . . . PPGPQG SEQ ID NO103 . . . IPGAPG SEQ ID NO117 . . . TSGHPG SEQ ID NO518 . . . PSGPPG SEQ ID NO483 . . . PPGPAG SEQ ID NO52 . . . FPGMKG SEQ ID NO409 . . . EKGPAG SEQ ID NO515 . . . MPGPRG SEQ ID NO523 . . . GIPGAP SEQ ID NO372 . . . NGDPGI SEQ ID NO471 . . . SPGPAG SEQ ID NO474 . . . PGPLGI SEQ ID NO477 . . . PGPPGT SEQ ID NO479 . . . APGLRG SEQ ID NO481 . . . GSPGPA SEQ ID NO484 . . . PGLMGA SEQ ID NO489 . . . GPPGPQ SEQ ID NO490 . . . GPAGIP SEQ ID NO441 . . . PPGPPG SEQ ID NO119 . . . GAPGLM SEQ ID NO498 . . . LPGAAG SEQ ID NO2188 . . . GPPGIN SEQ ID NO470 . . . PGLPGI SEQ ID NO504 . . . PGPKGD SEQ ID NO506 . . . GANGLP SEQ ID NO510 . . . GPPGIK SEQ ID NO512 . . . GAPGLR SEQ ID NO509 . . . GEVGPA SEQ ID NO514 . . . GKPGAN SEQ ID NO537 . . . PGAAGF SEQ ID NO539 . . . GDAGAP SEQ ID NO542 . . . GPAGPR SEQ ID NO545 . . . GGKGER SEQ ID NO548 . . . GPAGAN SEQ ID NO550 . . . VKGESG SEQ ID NO380 . . . TGPRGP SEQ ID NO177 . . . EPGPRG SEQ ID NO516 . . . GAPGPA SEQ ID NO519 . . . GTSGHP SEQ ID NO522 . . . GAPGLK SEQ ID NO525 . . . GEPGPR SEQ ID NO500 . . . PGPKGN SEQ ID NO527 . . . PPGAPG SEQ ID NO517 . . . TPGLQG SEQ ID 520 . . . SPGPQG SEQ ID NO472 . . . PGPQGV SEQ ID NO475 . . . AAGTPG SEQ ID NO478 . . . GNRGER SEQ ID NO480 . . . HPGSPG SEQ ID NO482 . . . GPAGPP SEQ ID NO485 . . . QGPPGP SEQ ID NO487 . . . GFPGMK SEQ ID NO493 . . . FPGAPG SEQ ID NO494 . . . FPGARG SEQ ID NO407 . . . GAIGPS SEQ ID NO495 . . . APGPLG SEQ ID NO2189 . . . IPGQPG SEQ ID NO501 . . . GAAGIK SEQ ID NO505 . . . GPPGVA SEQ ID NO158 . . . GPPGPS SEQ ID NO511 . . . TAGFPG SEQ ID NO513 . . . GPQGVK SEQ ID NO524 . . . GSPGPQ SEQ ID NO533 . . . QPGPPG SEQ ID NO536 . . . PGANGL SEQ ID NO529 . . . GPAGER SEQ ID NO543 . . . RGFDGR SEQ ID NO546 . . . APGLMG SEQ ID NO549 . . . PQGVKG SEQ ID NO541 . . . TGERGA SEQ ID NO540 . . . GSPGYQ SEQ ID NO526 . . . GAAGAR SEQ ID NO528 . . . TGAPGS SEQ ID NO502 . . . GTGGPP SEQ ID NO530 . . . GITGAR SEQ ID NO430 . . . GIAGPR SEQ ID NO535 . . . GLSGER SEQ ID NO387 . . . EGGPPG SEQ ID NO532 . . . GFPGAR SEQ ID NO408


6. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of collagen type I, said peptides comprising an N-terminal sequence selected from the group consisting of: Collagen I, alpha1 ISVPGP . . . SEQ ID NO23 FQGPPG . . . SEQ ID NO27 LDGAKG . . . SEQ ID NO31 VRGEPG . . . SEQ ID NO33 GGPPGP . . . SEQ ID NO35 DGVAGP . . . SEQ ID NO37 LTGSPG . . . SEQ ID NO39 RGVPGP . . . SEQ ID NO41 ERGEQG . . . SEQ ID NO43 PGERGV . . . SEQ ID NO45 ARGAPG . . . SEQ ID NO47 AKGDAG . . . SEQ ID NO49 AAGRVG . . . SEQ ID NO51 GADGPA . . . SEQ ID NO53 GQRGVV . . . SEQ ID NO55 GLPGQR . . . SEQ ID NO57 PMGPPG . . . SEQ ID NO59 DKGETG . . . SEQ ID NO61 SAGAPG . . . SEQ ID NO63 FDFSF . . . SEQ ID NO65 LPGPGG . . . SEQ ID NO26 SGLDGA . . . SEQ ID NO30 AKGEAG . . . SEQ ID NO68 IAGAPG . . . SEQ ID NO70 LPGPPG . . . SEQ ID NO72 AGPKGS . . . SEQ ID NO73 VPGPMG . . . SEQ ID NO24 KNGDDG . . . SEQ ID NO28 LPGERG . . . SEQ ID NO32 PGAKGA . . . SEQ ID NO34 NSGEPG . . . SEQ ID NO36 ERGSPG . . . SEQ ID NO38 QDGRPG . . . SEQ ID NO40 VGPAGK . . . SEQ ID NO42 RGEQGP . . . SEQ ID NO44 ANGAPG . . . SEQ ID NO46 PGDRGE . . . SEQ ID NO48 PIGNVG . . . SEQ ID NO50 PPGPAG . . . SEQ ID NO52 GPQGIA . . . SEQ ID NO54 QRGVVG . . . SEQ ID NO56 PGLPGP . . . SEQ ID NO58 MGPPGL . . . SEQ ID NO60 LQGPPG . . . SEQ ID NO62 RTGDAG . . . SEQ ID NO64 DFSF . . . SEQ ID NO66 QAGVMG . . . SEQ ID NO76 PAGERG . . . SEQ ID NO79 ARGERG . . . SEQ ID NO82 LTGPIG . . . SEQ ID NO85 AGPPGA . . . SEQ ID NO88 ATGFPG . . . SEQ ID NO91 GPPGPA . . . SEQ ID NO94 PGPMGP . . . SEQ ID NO25 ARGLPG . . . SEQ ID NO29 ATGAAG . . . SEQ ID NO67 GIAGAP . . . SEQ ID NO69 VQGPPG . . . SEQ ID NO71 RGSPGP . . . SEQ ID NO74 ARGQAG . . . SEQ ID NO77 KDGEAG . . . SEQ ID NO80 QGLPGP . . . SEQ ID NO83 AGLPGP . . . SEQ ID NO86 FAGPPG . . . SEQ ID NO75 NVGAPG . . . SEQ ID NO78 GEVGPP . . . SEQ ID NO81 IAGQRG . . . SEQ ID NO84 RGVVGL . . . SEQ ID NO87 EPGKQG . . . SEQ ID NO90 LAGPPG . . . SEQ ID NO89 PSGASG . . . SEQ ID NO92 VGPPGP . . . SEQ ID NO99 AGQRGV . . . SEQ ID NO95 VVGLPG . . . SEQ ID NO98 GKQGPS . . . SEQ ID NO93 ARGPAG . . . SEQ ID NO96 ASGPAG . . . SEQ ID NO97 GPPGPP . . . SEQ ID NO100


7. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of collagen type I, said peptides comprising an C-terminal sequence selected from the group consisting of: Collagen I, alpha1 . . . QLSYGY SEQ ID NO 101 . . . EKSTGG SEQ ID NO 102 . . . PPGPQG SEQ ID NO 103 . . . KGHRGF SEQ ID NO 104 . . . PSGPRG SEQ ID NO 105 . . . APGPQG SEQ ID NO 106 . . . APGPAG SEQ ID NO 107 . . . FPGAVG SEQ ID NO 108 . . . SEGPQG SEQ ID NO 109 . . . GANGAP SEQ ID NO 110 . . . ANGAPG SEQ ID NO 46 . . . SGPQGP SEQ ID NO 112 . . . EPGPVG SEQ ID NO 113 . . . EPGPTG SEQ ID NO 114 . . . RGFPGA SEQ ID NO 115 . . . KGPAGE SEQ ID NO 116 . . . RGSPGP SEQ ID NO 74 . . . LPGAKG SEQ ID NO 118 . . . AVGPAG SEQ ID NO 122 . . . PPGARG SEQ ID NO 120 . . . PGKAGE SEQ ID NO 121 . . . APGPDG SEQ ID NO 125 . . . PAGPAG SEQ ID NO 123 . . . AGPAGE SEQ ID NO 124 . . . KDGVRG SEQ ID NO 128 . . . RGERGF SEQ ID NO 126 . . . PAGPRG SEQ ID NO 127 . . . PGPAGF SEQ ID NO 131 . . . PAGPTG SEQ ID NO 129 . . . TGARGA SEQ ID NO 130 . . . SAGPPG SEQ ID NO 134 . . . EPGDAG SEQ ID NO 132 . . . PAGPPG SEQ ID NO 133 . . . GEVGPP SEQ ID NO 81 . . . ATGFPG SEQ ID NO 91 . . . NAGPPG SEQ ID NO 136 . . . PGPQGI SEQ ID NO 140 . . . GEKGSP SEQ ID 138 . . . GAPGTP SEQ ID NO 139 . . . IAGQRG SEQ ID NO 84 . . . GPQGIA SEQ ID NO 54 . . . PQGIAG SEQ ID NO 142 . . . GPSGEP SEQ ID NO 146 . . . GQRGVV SEQ ID NO 55 . . . RGERGF SEQ ID NO 126 . . . PVGPVG SEQ ID NO 149 . . . ERGPPG SEQ ID NO 147 . . . RGPPGP SEQ ID NO 148 . . . EQGPSG SEQ ID NO 152 . . . PQGPRG SEQ ID 150 . . . HRGFSG SEQ ID NO 151 . . . GPPGPP SEQ ID NO 100 . . . PRGPPG SEQ ID NO 153 . . . PPGPRG SEQ ID NO 154 . . . PPGPPG SEQ ID NO 119 . . . GPPSAG SEQ ID NO 156 . . . PPSAGF SEQ ID NO 157 . . . QMGPRG SEQ ID NO 161 . . . PPGPAG SEQ ID NO 52 . . . TPGPQG SEQ ID NO 160 . . . PGADGQ SEQ ID NO 164 . . . PGPPGA SEQ ID 162 . . . QGIAGQ SEQ ID 163 . . . PPGPKG SEQ ID NO 167 . . . AGSPGF SEQ ID NO 165 . . . LPGPSG SEQ ID NO 166 . . . PKGPAG SEQ ID NO 170 . . . PGERGA SEQ ID NO 168 . . . PMGPPG SEQ ID NO 59 . . . GPAGRP SEQ ID NO 173 . . . PPGPIG SEQ ID NO 174 . . . SPGEQG SEQ ID NO 172 . . . TGDAGP SEQ ID NO 175


8. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of collagen type IV, said peptides comprising an N-terminal sequence selected from the group consisting of: Collagen type IV IDGYRG . . . SEQ ID NO 609 MGPPGT . . . SEQ ID NO 610 DGLPGS . . . SEQ ID NO 611 PGSKGE . . . SEQ ID NO 612 RGFPGP . . . SEQ ID NO 613 PGPPGL . . . SEQ ID NO 614 GPPGPP . . . SEQ ID NO 100 GLPGSM . . . SEQ ID NO 615 GLPGQQ . . . SEQ ID NO 616 PGIGVQ . . . SEQ ID NO 618 GIGPPG . . . SEQ ID NO 611 PGLPGI . . . SEQ ID NO 504 PGPKGF . . . SEQ ID NO 621 SPGIPG . . . SEQ ID NO 619 PGMQGE . . . SEQ ID NO 620 LGSKGE . . . SEQ ID NO 617 KGQPGL . . . SEQ ID NO 622 RGPPGP . . . SEQ ID NO 148 IRGEPG . . . SEQ ID NO 626 FPGPPG . . . SEQ ID NO 627 GPLGEK . . . SEQ ID NO 625 DGVIGM . . . SEQ ID NO 629 PGNPGI . . . SEQ ID NO 630 PGLKGD . . . SEQ ID NO 624 IKGDKG . . . SEQ ID NO 631 PGSPGC . . . SEQ ID NO 632 PPSDEI . . . SEQ ID NO 623 GPPGVP . . . SEQ ID NO 633 DQGDQG . . . SEQ ID NO 634 GAPGPQ . . . SEQ ID NO 445 KGSIGI . . . SEQ ID NO 635 GIPGAP . . . SEQ ID NO 372 DRGPQG . . . SEQ ID NO 442 ERGSPG . . . SEQ ID NO 38 LQGIRG . . . SEQ ID NO 628 DGGVPN . . . SEQ ID NO 638 SGRDGL . . . SEQ ID NO 639 PGPMGP . . . SEQ ID NO 25 GPPGLM . . . SEQ ID NO 643 DGYRGP . . . SEQ ID NO 642 AEGLPG . . . SEQ ID NO 641 LPGFAG . . . SEQ ID NO 644 GIPGMP . . . SEQ ID NO 645 PPGRLG . . . SEQ ID NO 636 GPPGEK . . . SEQ ID NO 640 EKGQKG . . . SEQ ID NO 637 . . . LPGPDG SEQ ID NO 2214 . . . PGIPGT SEQ ID NO 2227 . . . ILGHVP SEQ ID NO 2212 . . . LRGIPG SEQ ID NO 760 . . . PGDIVF SEQ ID NO 2215 . . . PGLPGQ SEQ ID NO 2213 . . . PGFPGA SEQ ID NO 2218 . . . GNKGDP SEQ ID NO 2216 . . . SGYPGN SEQ ID NO 2217 . . . QQGNRG SEQ ID NO 2221 . . . PGPRGK SEQ ID NO 2219 . . . VSGPPG SEQ ID NO 2220 . . . VGQPGP SEQ ID NO 2222 . . . PGPPGP SEQ ID NO 458 . . . KRGPPG SEQ ID NO 2223 . . . GEPGMQ SEQ ID NO 2224 . . . SKGEKG SEQ ID NO 2226 . . . LHGFPG SEQ ID NO 2225 . . . GEPGPP SEQ ID NO 675


9. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of collagen type IV, said peptides comprising an C-terminal sequence selected from the group consisting of: Collagen type IV . . . RGPPGP SEQ ID NO 148 . . . SVDHGF SEQ ID NO 646 . . . PSVDHG SEQ ID NO 647 . . . VDHGFL SEQ ID NO 648 . . . PGQPGY SEQ ID NO 649 . . . QPGYTN SEQ ID NO 650 . . . PGLPGS SEQ ID NO 651 . . . GTPSVD SEQ ID NO 652 . . . SVGSPG SEQ ID NO 653 . . . LPGSMG SEQ ID NO 654 . . . GPPGVP SEQ ID NO 633 . . . PGFPGL SEQ ID NO 655 . . . PGLPGE SEQ ID NO 656 . . . GDPGPP SEQ ID NO 373 . . . HQGEMG SEQ ID NO 657 . . . GPPGLV SEQ ID NO 658 . . . PGIPGP SEQ ID NO 659 . . . PGFPGT SEQ ID NO 660 . . . PGLPGP SEQ ID NO 58 . . . DGIPGP SEQ ID NO 661 . . . SGPKGY SEQ ID NO 662 . . . GIGPPG SEQ ID NO 611 . . . PGPRGE SEQ ID NO 663 . . . GQGPPG SEQ ID NO 664 . . . PGAIGP SEQ ID NO 668 . . . KVDMGS SEQ ID NO 665 . . . PGIDGV SEQ ID NO 666 . . . GPKGPP SEQ ID NO 671 . . . PPGPPG SEQ ID NO 119 . . . KGHMGE SEQ ID NO 667 . . . GPKGLP SEQ ID NO 670 . . . SPGPPG SEQ ID NO 672 . . . KGLPGP SEQ ID NO 669 . . . GSVGYP SEQ ID NO 673 . . . PQGPPG SEQ ID NO 389 . . . PPGSPG SEQ ID NO 674 . . . GEPGPP SEQ ID NO 675 . . . AGNPGP SEQ ID NO 676 . . . FPGPQG SEQ ID NO 679 . . . PGYTNG SEQ ID NO 678 . . . SVDHGF SEQ ID NO 646 . . . LSGPPG SEQ ID NO 680 . . . PGPQGP SEQ ID NO 453 . . . GQGPPG SEQ ID NO 664 . . . PGAPGL SEQ ID NO 467 . . . GVMGTP SEQ ID NO 681 . . . PGIKGS SEQ ID NO 677 . . . PGPPGP SEQ ID NO 458 . . . GVSGPK SEQ ID NO 682 HVPGML . . . SEQ ID NO 2228 LPVPGQ . . . SEQ ID NO 2229 LGPPGL . . . SEQ ID NO 2230 GVPGQA . . . SEQ ID NO 2231 VPGQAQ . . . SEQ ID NO 2232 GPDGFL . . . SEQ ID NO 2233 QEGPLG . . . SEQ ID NO 2234 LPGEVL . . . SEQ ID NO 2235 RGIPGF . . . SEQ ID NO 2236 NRGLGF . . . SEQ ID NO 2238 IPSDTL . . . SEQ ID NO 2239 PAGEKG . . . SEQ ID NO 2240 GEKGNK . . . SEQ ID NO 2241 PVGPPG . . . SEQ ID NO 2242 DIVFRK . . . SEQ ID NO 2243 PPGPKG . . . SEQ ID NO 167 RGKPGM . . . SEQ ID NO 2244 PGTRGL . . . SEQ ID NO 2245 GEKGSK . . . SEQ ID NO 2246 EKGSKG . . . SEQ ID NO 2247 SGQPGL . . . SEQ ID NO 2248 AGIPQK . . . SEQ ID NO 2237


10. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of biglycan, decorin, lumican, versican, or perlecan said peptides comprising an N-terminal sequence selected from the group consisting of biglycan, Biglycan SVPKEI . . . SEQ ID NO 949 GLKLNY . . . SEQ ID NO 950 RISEAK . . . SEQ ID NO 951 NSGFEP . . . SEQ ID NO 952 LKSVPK . . . SEQ ID NO 953 AIELED . . . SEQ ID NO 954 LRISEA . . . SEQ ID NO 958 QCSDLG . . . SEQ ID NO 955 EAKLTG . . . SEQ ID NO 956 LLDLQN . . . SEQ ID NO 961 LTGIPK . . . SEQ ID NO 959 LKAVPK . . . SEQ ID NO 960 IELEDL . . . SEQ ID NO 957 Decorin IVIELG . . . SEQ ID NO 962 DEASGI . . . SEQ ID NO 963 VNNKIS . . . SEQ ID NO 964 NGLNQM . . . SEQ ID NO 965 LHLDGN . . . SEQ ID NO 966 LILVNN . . . SEQ ID NO 967 KITEIK . . . SEQ ID NO 969 GLPPSL . . . SEQ ID NO 970 SNPVQY . . . SEQ ID NO 971 SSGIEN . . . SEQ ID NO 968 Versican LLASDA . . . SEQ ID NO 972 LATVGE . . . SEQ ID NO 973 ETTVLV . . . SEQ ID NO 974 ENQDAR . . . SEQ ID NO 975 NGFDQC . . . SEQ ID NO 976 SLTVVK . . . SEQ ID NO 977 Lumican SLEDLQ . . . SEQ ID NO 978 LKEDAV . . . SEQ ID NO 979 HLQHNR . . . SEQ ID NO 980 LQHNRL . . . SEQ ID NO 985 Perlecan SIEYSP . . . SEQ ID NO 981 LVNFTR . . . SEQ ID NO 982 VSEAVV . . . SEQ ID NO 983 EVSEAV . . . SEQ ID NO 984


11. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of biglycan, decorin, lumican, versican, or perlecan, said peptides comprising an C-terminal sequence selected from the group consisting of: Biglycan . . . NNDISE SEQ ID NO 986 . . . YWEVQP SEQ ID NO 987 . . . EDLLRY SEQ ID NO 988 . . . RISEAK SEQ ID NO 951 . . . KIQAIE SEQ ID NO 989 . . . PETLNE SEQ ID NO 990 . . . LRKDDF SEQ ID NO 991 . . . LLRYSK SEQ ID NO 992 . . . ELRKDD SEQ ID NO 993 . . . KDLPET SEQ ID NO 994 . . . DLLRYS SEQ ID NO 995 . . . AFDGLK SEQ ID NO 996 . . . LNELHL SEQ ID NO 997 Decorin . . . GTNPLK SEQ ID NO 998 . . . EVPDDR SEQ ID NO 999 . . . GAFTPL SEQ ID NO 1000 . . . SSGIEN SEQ ID NO 968 . . . RVDAAS SEQ ID NO 1001 . . . LVKLER SEQ ID 1002 . . . GMKKLS SEQ ID NO 1003 . . . KDGDFK SEQ ID NO 1004 . . . HLDGNK SEQ ID NO 1005 . . . QPSTFR SEQ ID NO 1006 . . . AFQGMK SEQ ID NO 1007 Versican . . . CDVMYG SEQ ID NO 1008 . . . NGFDQC SEQ ID NO 976 . . . QNGINK SEQ ID NO 1009 . . . IGQDYK SEQ ID NO 1010 Lumican . . . QLTHNK SEQ ID NO 1011 . . . VSAAFK 1012 . . . GLKSLE 1013 Perlecan . . . EDAGSR SEQ ID NO 1014 . . . EFREVS SEQ ID NO 1015 . . . VAQQDS SEQ ID NO 1016 . . . SAKEFR SEQ ID NO 1017 . . . LEPEYR SEQ ID NO 1018


12. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of C-reactive protein (CRP), said peptides comprising an N-terminal sequence selected from the group consisting of: CRP AFVFPK SEQ ID NO 1033 SFGGNF SEQ ID NO 1305 FGQTDM SEQ ID NO 1306 VSLKAP SEQ ID NO 1172 KAFVFP SEQ ID NO 1167 EVFTKP SEQ ID NO 1201 TDMSRK SEQ ID NO 1307 MSRKAF SEQ ID NO 1308 SRKAFV SEQ ID NO 1309 VFPKES SEQ ID NO 1310 FPKESD SEQ ID NO 1311 KESDTS SEQ ID NO 1312 LSSTRG SEQ ID NO 1313 SSTRGY SEQ ID NO 1314 STRGYS SEQ ID NO 1080 KRQDNE SEQ ID NO 1315 WSKDIG SEQ ID NO 1316 SKDIGY SEQ ID NO 1317 SIILGQ SEQ ID NO 1318 YLGGPF SEQ ID NO 1322 ILGQEQ SEQ ID NO 1320 IYLGGP SEQ ID NO 1321 KYEVQG SEQ ID NO 1294 LGGPFS SEQ ID NO 1323 ALKYEV SEQ ID NO 1133 YTELSS SEQ ID NO 1325 VQGEVF SEQ ID NO 1324 KPQLWP SEQ ID NO 1157 QEQDSF SEQ ID NO 1328 ILIEWS SEQ ID NO 1326 LVGDIG SEQ ID NO 1327 QDSFGG SEQ ID NO 1331 RGYSIF SEQ ID NO 1329 GAEASI SEQ ID NO 1330 DTSYVS SEQ ID NO 1334 TIYLGG SEQ ID NO 1332 EINTIY SEQ ID NO 1333 SPDEIN SEQ ID NO 1337 AEASII SEQ ID NO 1335 TSWESA SEQ ID NO 1336 FVLSPD SEQ ID NO 1340 GGPFSP SEQ ID NO 1338 YEVQGE SEQ ID NO 1339 IVEFWV SEQ ID NO 1343 LKKGYT SEQ ID NO 1341 IILGQE SEQ ID NO 1319 EVQGEV SEQ ID NO 1346 ESDTSY SEQ ID NO 1344 RKAFVF SEQ ID NO 1342 SLKAPL SEQ ID NO 1222 FVFPK SEQ ID NO 1059 SDTSYV SEQ ID NO 1347 SYATKR SEQ ID NO 1349 LKAPLT SEQ ID NO 1173 IFSYAT SEQ ID NO 1348 WVDGKP SEQ ID NO 1352 YATKRQ SEQ ID NO 1350 EFWVDG SEQ ID NO 1351 GQEQDS SEQ ID NO 1355 VDGKPR SEQ ID NO 1353 LGQEQD SEQ ID NO 1354 LNWRA SEQ ID NO 1127 QSLVGD SEQ ID NO 1356 SPNVLN SEQ ID NO 1357 GEVFTK SEQ ID NO 1359 LNWRAL SEQ ID NO 1128 QGEVFT SEQ ID NO 1358 VRKSLK SEQ ID NO 1205 VFTKPQ SEQ ID NO 1153 IFWSKD SEQ ID NO 1181 ATKRQD SEQ ID NO 1362 KKGYTV SEQ ID NO 1360 FSYATK SEQ ID NO 1361 NWRAL SEQ ID NO 1253 FWSKDI SEQ ID NO 1363 VLNWRA SEQ ID NO 1249 DFVLSP SEQ ID NO 1366 NWRALK SEQ ID NO 1364 GSQSLV SEQ ID NO 1365 INTIYL SEQ ID NO 1372 VLSPDE SEQ ID NO 1367 LKYEVQ SEQ ID NO 1134 WRALKY SEQ ID NO 1375 KSLKKG SEQ ID NO 1371 GNFEGS SEQ ID NO 1369 TKPQLW SEQ ID NO 1345 RALKYE SEQ ID NO 1373 FVFPKE SEQ ID NO 1060 KGYTVG SEQ ID NO 1296 GPFSPN SEQ ID NO 1376 FYTELS SEQ ID NO 1374 TKRQDN SEQ ID NO 1368 SLKKGY SEQ ID NO 1370


13. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of C-reactive protein (CRP), said peptides comprising an C-terminal sequence selected from the group consisting of: CRP AFVFPK SEQ ID NO 1033 KPQLWP SEQ ID NO 1157 PDEINT SEQ ID NO 1377 KESDTS SEQ ID NO 1312 DSFGGN SEQ ID NO 1378 VFTKPQ SEQ ID NO 1153 LTKPLK SEQ ID NO 1379 SDTSYV SEQ ID NO 1347 DTSYVS SEQ ID NO 1334 KDIGYS SEQ ID NO 1380 STRGYS SEQ ID NO 1080 YATKRQ SEQ ID NO 1350 GAEASI SEQ ID NO 1330 DGKPRV SEQ ID NO 1381 VDGKPR SEQ ID NO 1353 SPNVLN SEQ ID NO 1357 GPFSPN SEQ ID NO 1376 NFEGSQ SEQ ID NO 1382 ALKYEV SEQ ID NO 1133 YEVQGE SEQ ID NO 1339 LKYEVQ SEQ ID NO 1134 KAFVFP SEQ ID NO 1167 VSLKAP SEQ ID NO 1172 LKAPLT SEQ ID NO 1173 LKKGYT SEQ ID NO 1341 LGQEQD SEQ ID NO 1354 NVNMWD SEQ ID NO 1383 PRVRKS SEQ ID NO 1384 TVGSEI SEQ ID NO 1385 SRKAFV SEQ ID NO 1309 GYSFTV SEQ ID NO 1386 IILGQE SEQ ID NO 1319 EGSQSL SEQ ID NO 1387 INTIYL SEQ ID NO 1372 WSKDIG SEQ ID NO 1316 EQDSFG SEQ ID NO 1388 NVLNWR SEQ ID NO 1389 ASGIVE SEQ ID NO 1390 NTIYLG SEQ ID NO 1391 VGAEAS SEQ ID NO 1392 TDMSRK SEQ ID NO 1307 FVFPKE SEQ ID NO 1060 QTDMSR SEQ ID NO 1394 TSYVSL SEQ ID NO 1396 MSRKAF SEQ ID NO 1308 PKESDT SEQ ID NO 1395 QDNEIL SEQ ID NO 1398 ESDTSY SEQ ID NO 1344 DNEILI SEQ ID NO 1397 TVGAEA SEQ ID NO 1401 NEILIF SEQ ID NO 1399 YTVGAE SEQ ID NO 1400 GNFEGS SEQ ID NO 1369 DIGNVN SEQ ID NO 1404 FEGSQS SEQ ID NO 1403 LNWRAL SEQ ID NO 1128 NWRALK SEQ ID NO 1364 LNWRA SEQ ID NO 1127 EVFTKP SEQ ID NO 1201 VFTKPQ SEQ ID NO 1153 KYEVQG SEQ ID NO 1294 RQDNEI SEQ ID NO 1405 IFWSKD SEQ ID NO 1181 KRQDNE SEQ ID NO 1315 VRKSLK SEQ ID NO 1205 SYVSLK SEQ ID NO 1407 FTKPQL SEQ ID NO 1406 TKPLKA SEQ ID NO 1408 SIFSYA SEQ ID NO 1409 SLKAPL SEQ ID NO 1222 GNVNMW SEQ ID NO 1410 SQSLVG SEQ ID NO 1411 GSQSLV SEQ ID NO 1365 GGNFEG SEQ ID NO 1413 LVGDIG SEQ ID NO 1327 FGGNFE SEQ ID NO 1412 RALKYE SEQ ID NO 1373 KKGYTV SEQ ID NO 1360 VQGEVF SEQ ID NO 1324 GKPRVR SEQ ID NO 1414 PFSPNV SEQ ID NO 1402 DMSRKA SEQ ID NO 1415 EILIEW SEQ ID NO 1416 QGEVFT SEQ ID NO 1358 FPKESD SEQ ID NO 1311 IGYSFT SEQ ID NO 1417 SPDEIN SEQ ID NO 1337 APLTKP SEQ ID NO 1393 FWSKDI SEQ ID NO 1363


14. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of elastin, said peptides comprising an N-terminal sequence selected from the group consisting of: Elastin GVPGAI SEQ ID NO 1898 AIPGGV SEQ ID NO 1899 GVPGGV SEQ ID NO 1900 ALGGGA SEQ ID NO 1901 LGGGAL SEQ ID NO 1902 GGALGP SEQ ID NO 1903 PLKPVP SEQ ID NO 1904 LKPVPG SEQ ID NO 1905 GLAGAG SEQ ID NO 1906 GLGAGL SEQ ID NO 1907 LGAGLG SEQ ID NO 1908 AGLGAF SEQ ID NO 1909 LVPGGV SEQ ID NO 1910 VADAAA SEQ ID NO 1911 KAAKAG SEQ ID NO 1912 PVGYPG SEQ ID NO 1913 ARFPGV SEQ ID NO 1914 RFPGVG SEQ ID NO 1915 VGPFGG SEQ ID NO 1916 GPQPGV SEQ ID NO 1917 PQPGVP SEQ ID NO 1918 TTGKLP SEQ ID NO 1919 LPYGYG SEQ ID NO 1920 GYGPGG SEQ ID NO 1921 FGAGAA SEQ ID NO 1922 GVLPGV SEQ ID NO 1923 VLPGVG SEQ ID NO 1924 TPAAAA SEQ ID NO 1928 VPGAIP SEQ ID NO 1926 AIPGIG SEQ ID NO 1927 VGVPGA SEQ ID NO 1931 PAAAAA SEQ ID NO 1929 AAAAAA SEQ ID NO 1930 GIPVVP SEQ ID NO 1934 AVGPGV SEQ ID NO 1932 GVPGVG SEQ ID NO 1933 ARPGVG SEQ ID NO 1937 IPGAAV SEQ ID NO 1935 GAAVPG SEQ ID NO 1936 SVGGVP SEQ ID NO 1940 RPGVGV SEQ ID NO 1938 VGGIPT SEQ ID NO 1939 VGVAPG SEQ ID NO 1943 VGGVPG SEQ ID NO 1941 GVGTPA SEQ ID NO 1942 GVPVAP SEQ ID NO 1946 VAPGVG SEQ ID NO 1944 GVAPGV SEQ ID NO 1945 LGAGIP SEQ ID NO 1949 GAGIPG SEQ ID NO 1947 PGGVAA SEQ ID NO 1948 PGALAA SEQ ID NO 1952 AKYGAA SEQ ID NO 1953 AGIPGL SEQ ID NO 1925 ALGGVG SEQ ID NO 1955 LGGVGI SEQ ID NO 1956 YGAAVP SEQ ID NO 1954 AGPAAA SEQ ID NO 1958 GPAAAA SEQ ID NO 1959 GVGIPG SEQ ID NO 1957 GLGVPG SEQ ID NO 1961 LGGIPP SEQ ID NO 1962 FGLVGA SEQ ID NO 1960 PLGGVA SEQ ID NO 1964 LGGVAA SEQ ID NO 1965 LGGVLG SEQ ID NO 1963 GVGLPG SEQ ID NO 1967 VGLPGV SEQ ID NO 1968 GGVAAR SEQ ID NO 1966 VPGVPV SEQ ID NO 1970 VPGVGI SEQ ID NO 1971 LPGVYP SEQ ID NO 1969 APGVGV SEQ ID NO 1973 VPGGVA SEQ ID NO 1974 VGISPE SEQ ID NO 1972 VPGGVF SEQ ID NO 1979 YPTGTG SEQ ID NO 1977 YPGGVL SEQ ID NO 1975 LGPGGK SEQ ID NO 1982 GVFYPG SEQ ID NO 1980 VPGAGV SEQ ID NO 1978 LAGAGL SEQ ID NO 1985 GPGGKP SEQ ID NO 1983 VFYPGA SEQ ID NO 1981 LGAFPA SEQ ID NO 1988 AGAGLG SEQ ID NO 1986 PGGKPL SEQ ID NO 1984 LGVSAG SEQ ID NO 1991 AFPAVT SEQ ID NO 1989 GAGLGA SEQ ID NO 1987 FPGVGV SEQ ID NO 1994 VPGVGL SEQ ID NO 1992 AVTFPG SEQ ID NO 1990 PGVPLG SEQ ID NO 1996 KPGAPG SEQ ID NO 1995 PGVGLP SEQ ID NO 1993 YGPGGV SEQ ID NO 1999 GYPIKA SEQ ID NO 1997 PKAPGV SEQ ID NO 1493 GAGVPG SEQ ID NO 2002 AGYPTG SEQ ID NO 2000 PKLPGG SEQ ID NO 1998 IPGIGG SEQ ID NO 2005 AGVPGV SEQ ID NO 2003 TGVGPQ SEQ ID NO 2001 GPGFGP SEQ ID NO 1976 IGGIAG SEQ ID NO 2006 GVPGVP SEQ ID NO 2004 VPGVGV SEQ ID NO 2008 PGFGPG SEQ ID NO 1950 GIAGVG SEQ ID NO 2007 AVPGVV SEQ ID NO 2011 VPVGVA SEQ ID NO 2009 PGVVGV SEQ ID NO 1951 GVGAGG SEQ ID NO 2014 VPGVVS SEQ ID NO 2012 VPGAGI SEQ ID NO 2010 FGLVPG SEQ ID NO 2017 VGAGGF SEQ ID NO 2015 YGARPG SEQ ID NO 2013 PGVGLA SEQ ID NO 2020 GLVPGV SEQ ID NO 2018 VGVGGI SEQ ID NO 2016 LRAAAG SEQ ID NO 2023 GVGLAP SEQ ID NO 2021 LVPGVG SEQ ID NO 2019 GIPGLG SEQ ID NO 2026 LVGAAG SEQ ID NO 2024 VGLAPG SEQ ID NO 2022 VGIPGG SEQ ID NO 2032 LGVGVG SEQ ID NO 2027 LVPGGP SEQ ID NO 2025 GLVGAA SEQ ID NO 2035 AVPGVL SEQ ID NO 2030 VGVPGL SEQ ID NO 2028 GGVLGG SEQ ID NO 2038 IPGGVV SEQ ID NO 2033 VLGGLG SEQ ID NO 2031 GVAARP SEQ ID NO 2041 VGAAGL SEQ ID NO 2036 VVGAGP SEQ ID NO 2034 GVYPGG SEQ ID NO 2044 GAGQFP SEQ ID NO 2039 LGGLGV SEQ ID NO 2037 AAVPGV SEQ ID NO 2047 VAARPG SEQ ID NO 2042 AFQFPL SEQ ID NO 2041 VPGVLG SEQ ID NO 2050 AAGLGG SEQ ID NO 2048 RPGFGL SEQ ID NO 2043 ISPEAQ SEQ ID NO 2053 GQFPLG SEQ ID NO 2051 FGPGVV SEQ ID NO 2046 LGALGG SEQ ID NO 2056 GVLPGA SEQ ID NO 2054 FPGALV SEQ ID NO 2049 GKPLKP SEQ ID NO 2059 PQAAAA SEQ ID NO 2077 ALVPGG SEQ ID NO 2052 AGLGAG SEQ ID NO 2062 VPGVPG SEQ ID NO 2057 AAAGLG SEQ ID NO 2029 VTFPGA SEQ ID NO 2065 IAGVGT SEQ ID NO 2060 VGAGVP SEQ ID NO 2055 GLPGVY SEQ ID NO 2068 VVGVPG SEQ ID NO 2063 GGLGAL SEQ ID NO 2058 GAFAGI SEQ ID NO 2071 AGIPVV SEQ ID NO 2066 VGAGPA SEQ ID NO 2061 YTTGKL SEQ ID NO 2074 GARPGV SEQ ID NO 2069 LGVGGL SEQ ID NO 2064 PGVGVA SEQ ID NO 2075 VAGVPS SEQ ID NO 2072 FPLGGV SEQ ID NO 2067 LAPGVG SEQ ID NO 2078 VGTPAA SEQ ID NO 2076 PGFGLS SEQ ID NO 2070 LPYTTG SEQ ID NO 2045 GPGVVG SEQ ID NO 2073


15. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of elastin, said peptides comprising an C-terminal sequence selected from the group consisting of: Elastin PGGVPG SEQ ID NO 2079 PGAGLG SEQ ID NO 2080 GAGLGA SEQ ID NO 1987 GALGGG SEQ ID NO 2081 LGGGAL SEQ ID NO 1902 GGGALG SEQ ID NO 2082 GLGAFP SEQ ID NO 2083 LGAFPA SEQ ID NO 1988 LGAGLG SEQ ID NO 1908 VPGGVA SEQ ID NO 1974 ADAAAA SEQ ID NO 2084 VPTGAG SEQ ID NO 2087 RFPGVG SEQ ID NO 1915 VGVLPG SEQ ID NO 2086 VPLGYP SEQ ID NO 2089 PKAPGV SEQ ID NO 1493 GVGPFG SEQ ID NO 2088 IPGIGG SEQ ID NO 2005 LPYGYG SEQ ID NO 1920 AAAAAK SEQ ID NO 2090 IGGIAG SEQ ID NO 2006 GIAGVG SEQ ID NO 2007 AIPGIG SEQ ID NO 1927 GVPGVG SEQ ID NO 1933 AAAAKA SEQ ID NO 2091 VVGVPG SEQ ID NO 2063 VVSPEA SEQ ID NO 2094 GVGVPG SEQ ID NO 2092 PVVPGA SEQ ID NO 2093 GIPTYG SEQ ID NO 2096 VGGIPT SEQ ID NO 1939 GGIPTY SEQ ID NO 2095 GVGAGG SEQ ID NO 2014 GVGVGG SEQ ID NO 2097 GVGGIP SEQ ID NO 2098 VAPGVG SEQ ID NO 1944 GFPGFG SEQ ID NO 2099 FGVGVG SEQ ID NO 2100 APGVGL SEQ ID NO 2104 PGVGIS SEQ ID NO 2101 SPEAQA SEQ ID NO 2102 VAPGIG SEQ ID NO 2107 PGVGVA SEQ ID NO 2075 GVGVAP SEQ ID NO 2103 AAGLGA SEQ ID NO 2109 GIGPGG SEQ ID NO 2105 APGIGP SEQ ID NO 2106 AGVPGL SEQ ID NO 2112 RAAAGL SEQ ID NO 2108 AAAGLG SEQ ID NO 2029 GAGPAA SEQ ID NO 2113 GVPGLG SEQ ID NO 2110 GVPGFG SEQ ID NO 2111 GLGGLG SEQ ID NO 2115 VLGGLG SEQ ID NO 2031 VGIPGG SEQ ID NO 2032 PAAAAK SEQ ID NO 2117 PAAAAA SEQ ID NO 1929 VPGVGG SEQ ID NO 2114 AARPGF SEQ ID NO 2118 GLGVPG SEQ ID NO 1961 PGVGGL SEQ ID NO 2116 GPGIPG SEQ ID NO 2121 RPGFGL SEQ ID NO 2043 GVAARP SEQ ID NO 2041 ALGPGG SEQ ID NO 2124 LSPIFP SEQ ID NO 2119 AQAAAA SEQ ID NO 2120 KPVPGG SEQ ID NO 2127 GPGGVA SEQ ID NO 2122 LGALGG SEQ ID NO 2056 VTFPGA SEQ ID NO 2065 GLGALG SEQ ID NO 2123 PGAIPG SEQ ID NO 2146 VPQPGA SEQ ID NO 2130 GALGPG SEQ ID NO 2125 TPAAAA SEQ ID NO 1928 AGVKPK SEQ ID NO 2133 AFPAVT SEQ ID NO 1989 AVTFPG SEQ ID NO 1990 YGYGPG SEQ ID NO 2136 AAAAYK SEQ ID NO 2128 AAKAGA SEQ ID NO 2129 GVGTPA SEQ ID NO 1942 PGVPTG SEQ ID NO 2131 GVPTGA SEQ ID NO 2132 AAAAAA SEQ ID NO 1930 PIKAPK SEQ ID NO 2134 KLPGGY SEQ ID NO 2135 GVPGAG SEQ ID NO 2140 VGTPAA SEQ ID NO 2076 VPGVPG SEQ ID NO 2057 ARPGVG SEQ ID NO 1937 GIGGIA SEQ ID NO 2137 GTPAAA SEQ ID NO 2138 YGVGAG SEQ ID NO 2144 IPVVPG SEQ ID NO 2139 VGVPGA SEQ ID NO 1931 VGAGGF SEQ ID NO 2015 GAGIPG SEQ ID NO 1947 PEAAAK SEQ ID NO 2141 GISPEA SEQ ID NO 2149 IPTYGV SEQ ID NO 2145 TYGVGA SEQ ID NO 2143 VPGAPG SEQ ID NO 2150 AGGFPG SEQ ID NO 2147 PTYGVG SEQ ID NO 2142 APGVGV SEQ ID NO 1973 VGVAPG SEQ ID NO 1943 GIPGVA SEQ ID NO 2148 PGIGPG SEQ ID NO 2152 PGVGLA SEQ ID NO 2020 VGLAPG SEQ ID NO 2022 LGVGVG SEQ ID NO 2027 GAGVPG SEQ ID NO 2002 LAPGVG SEQ ID NO 2078 LGGLGA SEQ ID NO 2157 PGVGVG SEQ ID NO 2169 GGVAAA SEQ ID NO 2151 GPAAAA SEQ ID NO 1959 GLGVGG SEQ ID NO 2153 PGLGVG SEQ ID NO 2154 GVGGLG SEQ ID NO 2159 GLGVGA SEQ ID NO 2155 ALAAAK SEQ ID NO 2156 VAARPG SEQ ID NO 2042 VGAGPA SEQ ID NO 2061 AGPAAA SEQ ID NO 1958 IFPGGA SEQ ID NO 2163 GGLGVG SEQ ID NO 2158 LGVGGL SEQ ID NO 2064 AAKAAK SEQ ID NO 2166 AGQFPL SEQ ID NO 2160 GGVAAR SEQ ID NO 1966 PGGVAA SEQ ID NO 1948 GFGLSP SEQ ID NO 2161 PIFPGG SEQ ID NO 2162 VGVPGL SEQ ID NO 2028 AAKFGA SEQ ID NO 2164 AAKYGA SEQ ID NO 2165 PGVLGG SEQ ID NO 2085 AGLGAL SEQ ID NO 2167 RPGVGV SEQ ID NO 1938 GIPGGV SEQ ID NO 2168 LKPVPG SEQ ID NO 1905 GGFPGF SEQ ID NO 2173 IPPAAA SEQ ID NO 2170 ALVPGG SEQ ID NO 2052 VGGVPG SEQ ID NO 1941 ARPGFG SEQ ID NO 2171 VLPGAR SEQ ID NO 2172 GVAPGV SEQ ID NO 1945 GLSPIF SEQ ID NO 2174 PTGAGV SEQ ID NO 2175 VAPVGV SEQ ID NO 2126 GIPVVP SEQ ID NO 1934 AGAAGK SEQ ID NO 2176


16. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of collagen type V, said peptides comprising an N-terminal sequence selected from the group consisting of: Collagen type V GDPGPP SEQ ID NO 373 LRGIPG SEQ ID NO 760 IGPPGI SEQ ID NO 761 LQGPPG SEQ ID NO 62 IGSLGH SEQ ID NO 762 IRGPPG SEQ ID NO 763 ANGSPG SEQ ID NO 764 LIGTPG SEQ ID NO 765 LPGEPG SEQ ID NO 766 IPGRPG SEQ ID NO 767 GPDGPP SEQ ID NO 768 QPGPSG SEQ ID NO 769 LKGNEG SEQ ID NO 770 ERGHPG SEQ ID NO 771 GPPGEQ SEQ ID NO 772 FPGPKG SEQ ID NO 491 PFRFGG SEQ ID NO 773 ESQAQA SEQ ID NO 774 LLGPKG SEQ ID NO 775 QQGNPG SEQ ID NO 776 KEGPPG SEQ ID NO 777 IGLIGP SEQ ID NO 778 GPPGPP SEQ ID NO 100 LGPPGE SEQ ID NO 779 GPPGPK SEQ ID NO 780 SLGHPG SEQ ID NO 781 KDGIPG SEQ ID NO 782 PVGEPG SEQ ID NO 783 LRGIPG SEQ ID NO 760 KDGIPG SEQ ID NO 782 ANGSPG SEQ ID NO 764 LIGTPG SEQ ID NO 765 PGEPGP SEQ ID NO 784 KDGIP SEQ ID NO 813 VLGPQG SEQ ID NO 814 DGIPGP SEQ ID NO 661 QMGPPG SEQ ID NO 802 LLGAPG SEQ ID NO 785 GLPGLE SEQ ID NO 786 ALRGPA SEQ ID NO 810 LALRGP SEQ ID NO 788 LTGRPG SEQ ID NO 789 ETGFQG SEQ ID NO 808 LRGFPG SEQ ID NO 790 KTGPIG SEQ ID NO 791 EAGHPG SEQ ID NO 811 PGPKGN SEQ ID NO 527 EQGLPG SEQ ID NO 793 GSKGPM SEQ ID NO 809 AIGGPP SEQ ID NO 794 GPNGDP SEQ ID NO 795 EKGHPG SEQ ID NO 812 LLGPRG SEQ ID NO 797 GPDGPP SEQ ID NO 768 GPKGSI SEQ ID NO 816 GPKGDP SEQ ID NO 799 GDPGPP SEQ ID NO 373 ELGFQG SEQ ID NO 817 LQGPPG SEQ ID NO 62 EKGHIG SEQ ID NO 801 LRGPAG SEQ ID NO 818 SRGERG SEQ ID NO 803 EKGKSG SEQ ID NO 804 EDGERG SEQ ID NO 815 LPGEPG SEQ ID NO 766 PQGAIG SEQ ID NO 806 GIPGEK SEQ ID NO 787 PPGRPG SEQ ID NO 792 PGPPGE SEQ ID NO 796 LLGPKG SEQ ID NO 775 GPPGEK SEQ ID NO 640 ERGPNG SEQ ID NO 798 GPMGLT SEQ ID NO 807 GPIGEK SEQ ID NO 805 PGIPGE SEQ ID NO 800 QMGPPG SEQ ID NO 802


17. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of collagen type V, said peptides comprising an C-terminal sequence selected from the group consisting of: Collagen type V PIGSLG SEQ ID NO 819 PVGEPG SEQ ID NO 783 PGPPGE SEQ ID NO 796 PPGPTG SEQ ID NO 820 PKGPPG SEQ ID NO 821 VAGPLG SEQ ID NO 822 RPGVTG SEQ ID NO 823 MMPFQF SEQ ID NO 824 PGAAGP SEQ ID NO 825 PVGPAG SEQ ID NO 826 HEGPTG SEQ ID NO 827 EPGPRG SEQ ID NO 516 GPPGLP SEQ ID NO 828 GQGPPG SEQ ID NO 664 PPGPPG SEQ ID NO 119 AGSPGE SEQ ID NO 829 PVGALG SEQ ID NO 830 EQGLPG SEQ ID NO 793 YPGPRG SEQ ID NO 831 HPGQRG SEQ ID NO 832 GGGGDA SEQ ID NO 833 LIGPPG SEQ ID NO 834 GLPGLK SEQ ID NO 835 PPGPPG SEQ ID NO 119 PPGPIG SEQ ID NO 174 KGLPGL SEQ ID NO 836 PGPPGP SEQ ID NO 458 QMGPPG SEQ ID NO 802 LLGAPG SEQ ID NO 785 RPGVTG SEQ ID NO 823 QQARLA SEQ ID NO 837 PPGHPG SEQ ID NO 838 PQGPRG SEQ ID NO 150 PLGPPG SEQ ID NO 839 GEPGLL SEQ ID NO 840 EVGPLG SEQ ID NO 841 IMMPFQ SEQ ID NO 842 GLPGLK SEQ ID NO 835 PLGPSG SEQ ID NO 843 AAGPSG SEQ ID NO 844 PPGPLG SEQ ID NO 845 PPGSRG SEQ ID NO 846 PAGPMG SEQ ID NO 847 PPGSGG SEQ ID NO 848 PPGPPG SEQ ID NO 119 GLPGPV SEQ ID NO 849 LPGPVG SEQ ID NO 850 KPGPDG SEQ ID NO 851 LPGPQG SEQ ID NO 852 VIQPLP SEQ ID NO 853 RGPNGP SEQ ID NO 854 PGPQGS SEQ ID NO 855 EKGPLG SEQ ID NO 856 PLGPPG SEQ ID NO 839 PPGPLG SEQ ID NO 845 TGPKGE SEQ ID NO 858 GPKGSI SEQ ID NO 816 GPPGAA SEQ ID NO 857 PPGPAG SEQ ID NO 52 PPGPTG SEQ ID NO 820 LLGAPG SEQ ID NO 785 PLGPLG SEQ ID NO 863 PPGSRG SEQ ID NO 846 PSGLPG SEQ ID NO 860 LPGPPG SEQ ID NO 72 KPGPRG SEQ ID NO 862 PPGPAG SEQ ID NO 52 PPGLPG SEQ ID NO 861 PKGPPG SEQ ID NO 821 QGLPGL SEQ ID NO 859 PPGSRG SEQ ID NO 846


18. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by an N-terminal amino acid sequence present in peptides produced by cleavage of collagen type VI, said peptides comprising an N-terminal sequence selected from the group consisting of: Collagen type VI YRGPEG SEQ ID NO 883 PIGPKG SEQ ID NO 865 GIGIGN SEQ ID NO 885 ISGPRG SEQ ID NO 886 PGPAGP SEQ ID NO 887 VAAKPA SEQ ID NO 888 GEPGPP SEQ ID NO 675 RGPIGS SEQ ID NO 889 PPPPQP SEQ ID NO 890 AQGPAG SEQ ID NO 891 LIGEQG SEQ ID NO 892 PGLIGE SEQ ID NO 893 GEPGLN SEQ ID NO 894 IGPKGI SEQ ID NO 895 VAVVQH SEQ ID NO 896 FGPSAA SEQ ID NO 897 GPKGET SEQ ID NO 898 LGPMGV SEQ ID NO 899 PGEPGP SEQ ID NO 784


19. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of collagen type VI, said peptides comprising an C-terminal sequence selected from the group consisting of: Collagen type VI GDEGPP SEQ ID NO 900 GNADIT SEQ ID NO 901 PAGPPG SEQ ID NO 133 DPGLMG SEQ ID NO 902 PEVPRP SEQ ID NO 903 TGPKGI SEQ ID NO 904 GDEGGP SEQ ID NO 905 PARSAS SEQ ID NO 906 TPAPPG SEQ ID NO 915 ISGPRG SEQ ID NO 886 GISGPR SEQ ID NO 907 GIGNRG SEQ ID NO 908 YRGYPG SEQ ID NO 909 KVEFSL SEQ ID NO 910 GVPGRD SEQ ID NO 911 PGETGK SEQ ID NO 912 RTGPLG SEQ ID NO 913 APGERG SEQ ID NO 914


20. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a N-terminal amino acid sequence present in peptides produced by cleavage of vimentin, said peptides comprising an N-terminal sequence selected from the group consisting of: Vimentin LLQDSV SEQ ID NO 2182 FADLSE SEQ ID NO 2183 ISLPLP SEQ ID NO 2184


21. A method as claimed in claim 1, wherein said immunological binding partner specifically binds a neo-epitope constituted by a C-terminal amino acid sequence present in peptides produced by cleavage of vimentin, said peptides comprising an C-terminal sequence selected from the group consisting of: Vimentin SVPGVR SEQ ID NO 2185 SVPGVL SEQ ID NO 2186


22. A method as claimed in claim 1, subject to the further proviso that if the neo-epitopes are formed by cleavage of type I collagen, they are not at locations where type I collagen is cleaved by trypsin.
 23. A method of immunoassay to measure neo-epitope containing protein fragments naturally present in body fluid sample, wherein said immunoassay is conducted by a method comprising: contacting protein fragments naturally present in said sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to said immunological binding partner to measure therein protein fragments comprising said neo-epitope, and wherein said protein is neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, elastin, collagen type I, collagen type IV, collagen type V or collagen type VI, CRP, or vimentin, subject to the proviso that when the neo-epitopes are formed by cleavage of type I collagen, the cleavage is not at a site at which collagen type I is cleaved by cathepsin K. 